Compositions and methods for treating cancer

ABSTRACT

Provided herein, inter alia, are compositions and methods useful for treating hyperproliferative diseases, including cancer and non-malignant hyperproliferative diseases.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a continuation application of U.S. patentapplication Ser. No. 14/505,288, filed Oct. 2, 2014, which is acontinuation application of international PCT application No.PCT/US2013/035010, filed Apr. 2, 2013, which claims the benefit of U.S.Provisional Patent Application No. 61/619,289, filed Apr. 2, 2012, whichare incorporated herein by reference in their entirety and for allpurposes.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

This invention was made with Government support under DAMD17-03-1-0381,awarded by the U.S. Army, Medical Research and Materiel Command. TheGovernment has certain rights in the invention.

BACKGROUND

Lung Cancer is the leading cause of cancer death in women and men in theU.S. It is estimated that there will be more than 230,000 patientsdiagnosed with lung cancer this year, with more than 160,000 deaths, andnon-small cell lung cancer (NSCLC) will account for more than threequarters of these cases (1). Survival rates of NSCLC, the predominanttype of lung cancer are unacceptably low, and new approaches to treatand prevent this disease are urgently needed. Feverfew extracts andparthenolide show anticancer activity in NSCLC in vitro, whileparthenolides modified to enhance solubility also show significantanticancer activity in vivo in NSCLC models. This anticancer effect maybe associated with the ability of these substances to inhibit thenuclear transcription factor kappaB (NFκB) possibly leading in turn tothe promotion of tumor cell apoptosis (3). Disclosed herein is a studyof the role of feverfew extract and parthenolide in the treatment andprevention of NSCLC.

At diagnosis, about 70% of breast cancer patients have tumors thatexpress estrogen receptors (ER) and/or progesterone receptors (PR).Patients with ER+ tumors can be treated with hormonal therapy such astamoxifen which was the first effective targeted therapy for breastcancer. However, all advanced ER+ tumors eventually develop endocrineresistance, and there is an urgent need for new interventions to stopendocrine resistance. Mechanisms of endocrine resistance include HER2overexpression (occurs in 15-20% of patients) and increased activationof NFκB signaling pathways. Activation of tumor NFκB occurs in ER+breast cancers with resistance to hormonal therapy and in HER2+ breasttumors with resistance to Trastuzumab (Herceptin). Parthenolideanalogues that block activation of NFκB are active in restoring breasttumor responses to hormonal and Trastuzumab therapies, therebypotentially helping to improve patient outcomes

Tuberous sclerosis complex (TSC) is an autosomal dominant disorder withdevelopment of hamartomatous lesions in many organs (71-74). Somelesions grow progressively and require clinical intervention. TSC is dueto mutations in either TSC1 or TSC2 genes. TSC1 (hamartin)/TSC2(tuberin) protein complexes serve a critical role in negativelyregulating mTOR complex 1 (mTORC1) which exerts downstream effects oncell transcription, translation, metabolism and proliferation. mTORC1 isconstitutively active in cells lacking either TSC1 or TSC2 and inhamartomas of TSC patients. TSC can occur in association with pulmonarylymphangioleiomyomatosis (LAM), a progressive and often fatalinterstitial lung disease that occurs largely in women and ischaracterized by proliferation of abnormal smooth muscle cells.TSC2-null smooth muscle cells in angiomyolipomas (AML) are very similarto those of pulmonary LAM, and data suggest LAM may be the result ofbenign cell metastases. Rapamycin and related drugs (everolimus) whichbind and inhibit mTORC1 have clinical activity for therapy of TSC,including renal AML and pulmonary LAM (72). However, rapamycin does notelicit complete TSC regression in most cases, and termination of therapyoft leads to lesion re-growth. New therapeutic approaches to control thegrowth of TSC-related proliferative diseases are urgently needed.

The poor prognosis of advanced non-small cell lung cancer (NSCLC) isdue, in part, to emergence of tumor resistance to chemotherapy (1).Recent data indicate that human tumors contain a small subset of cancerstem cells (CSC) responsible for drug resistance and tumor maintenance(2-5). If such minute subsets of CSC drive tumor formation and drugresistance, therapies targeting the bulk tumor mass but not CSC willfail. Since it is hypothesized that CSC are responsible for tumorregeneration after chemo-therapy (6), we performed preliminary studiesto determine if drug treatment could enrich for CSCs. Treatment of humanlung tumor NSCLC cell lines (A549, H23) with increasing doses ofcisplatin resulted in selection of drug-resistant cells (DRC)(6). UsingFluorescence Activated Cell Sorting (FACS), DRC were selected forexpression of cell surface CD133+ and cytosolic ALDH1 by Aldefluor assay(7,8), yielding cell subsets amounting to 2%-5% of bulk tumor cellpopulations of H23 and A549, respectively. Furthermore, CSC-like cellsgrew as tumor spheres, maintained self-renewal capacity anddifferentiated, with differentiated progenitors losing CD133 expressionand acquiring drug sensitivity. These initial studies suggest thatchemotherapy leads to propagation of CSC. Importantly, resistance ofCSC-like cells to cisplatin was fully reversed by treatment withparthenolide (PTL). The antitumor effect of PTL has been reported toselectively kill CSCs through inhibition of nuclear factor-κB (NF-κB)which is markedly activated by chemotherapy (9).

The aromatic plant known as feverfew (Tanacetum parthenium) has beenused in folk medicine to treat a number of different human maladies. Theplant is notably rich in a family of compounds known as sesquiterpenelactones, particularly parthenolide. One of the most importantcharacteristics of parthenolide is its antitumor activity, with reportsof cytotoxic activity against epidermoid carcinoma, fibrosarcoma,hepatocellular carcinoma, breast and prostate cancer, and leukemia andlymphoma cells. This compound has been demonstrated in vitro to inhibitdifferent type of cancer cells (2,4,5). Parthenolide may promoteapoptosis in neoplastic cells in part by inhibiting the cancer-promotingfactor, NFκB, a critical regulator of genes involved in tumor cellproliferation, antiapoptosis (anti-cell death), DNA damage responses andangiogenesis (2,4,6,7). Inhibition of NFκB signaling leads to reducedexpression of many proteins including antiapoptotic proteins. Thedownstream consequences of these effects lead to cell cycle arrest andcell death (2,8). Unfortunately, both feverfew herbal extract and nativeparthenolide have limited in vivo activity due to poor bioavailability(9). Hence, the inventors have developed improved derivatives of thesenaturally-occurring compounds to be used as drug-like agents to treatand prevent lung cancer and other human malignancies.

Breast cancer is a worldwide health concern with about 1,000,000 millionnew cases each year. In the clinic, endocrine therapy is an importantintervention in women with breast cancers that express estrogen receptor(ER), and treatment with tamoxifen has enhanced patient survival. Thesuccess of endocrine therapy is dependent on tight regulation of breastcell growth by steroids and growth factor receptors (27) most patientseventually stop responding to antiestrogen therapy. Resistance totamoxifen (TAM) and aromatase inhibitors represents a major drawback totreatment of hormone-dependent breast cancer, and new options forendocrine therapy are urgently needed to reverse this outcome. Emergingdata now confirm the existence of previously-unsuspected interactionsbetween growth factor and estrogen signaling pathways that contribute togrowth regulation in breast cancers. Targeting this signaling axis maypromote introduction of more effective and less toxic antihormonetreatments for human breast cancers (28-30).

Subversion of growth factor receptors often occurs in malignancy, andmembers of the HER family are often implicated in cancer (31). EGFreceptor (EGFR), a 170-kD transmembrane protein, has an extracellularligand-binding domain, a membrane-spanning region and a cytoplasmicEGF-stimulated tyrosine kinase. On EGF binding, EGFR undergoesdimerization and autophosphorylation on tyrosine residues. This resultsin activation of downstream protein kinases, such as MAP kinase and PI3kinase/Akt kinase, and subsequent stimulation of transcription factors.The HER family includes other receptor tyrosine kinases including HER-2,a 185-kD kinase encoded by HER-2/neu oncogene, as well as HER-3 andHER-4. HER-2 receptor functions similar to EGFR (30-33), and, on bindingof ligand to EGF, HER-3 or HER-4, HER-2 is often recruited as apreferred partner to form heterodimers that activate downstreamsignaling for growth and survival. Overexpression of HER-2 or relatedHER receptors occurs in two-thirds of sporadic breast tumors, whileHER-2 over-expression/amplification is found in 25-30% of breast cancers(32,33). Overexpression of HER-2 is generally a marker of poor prognosis(7), and it associates with failure of antiestrogen therapy in theclinic (28,34-38).

It is generally held that biologic activity of estrogen in breast cellsis mediated by its binding with high-affinity ER in the nucleus (27,39)(see FIG. 10).

Upon estrogen binding, ER undergoes an activating conformational changethat allows association with coactivators and target genes, thuspromoting regulation of gene transcription. Ligand-bound ER in thenucleus functions as a transcription factor. Blocking estrogen bindingto ER is the basis of the action of tamoxifen, a partial agonist thatlimits proliferative effects of estrogen in breast, while fulvestrant(Faslodex) is a new generation antiestrogen with a mechanism of actionleading to downregulation of ER in tumors (42,43). In addition tonuclear actions of estrogens, numerous reports document rapid effects ofestradiol mediated by a membrane-associated ER that derives from thesame transcript as nuclear ER (34-46) (FIG. 10). Interactions between ERand growth factor receptors occur in lipid rafts, assemblies ofcholesterol and sphingolipids in plasma membrane (47). Caveolae arespecialized rafts present in most cells (48,49), but they are markedlyreduced or absent in breast cancer cells (50). Caveolae and lipid raftsare enriched in growth factor receptors, including HER-2 receptors (48),and a portion of ER also localize in caveolae and lipid rafts (51,52).

Conversion of estrogen-sensitive to -resistant tumors after the start ofantiestrogen therapy is a major problem in the clinic (65). Thisresistance may be due, in part, to enhanced growth factor signaling, acellular response to antiestrogen treatment that eventually results inincreased phosphorylation of ER and/or coactivators, such as AIB1(54).Convergence between growth factor and estrogen signaling may elicit asynergistic feed-forward circuit with more robust cell growth (FIG. 10).It is notable that tumors with HER-2 overexpression tend to respondpoorly to antiestrogens (69-72). A meta-analysis of 7 clinical trialsindicated that breast tumors with HER-2 overexpression were resistant totamoxifen (73-75). Such clinical data offer further evidence of thebiologic interaction between HER-2 and ER. To evaluate the relation ofHER-2 to clinical antiestrogen resistance, we used estrogen-responsive,MCF-7 cells with defined levels of ER that were transfected with HER-2gene (MCF-7/HER-2). MCF-7 cells+HER-2 overexpression and parent MCF-7cells were grown as xenografts in nude mice as before (28) (FIG. 11). Asexpected, growth of parent cells was markedly suppressed by tamoxifenand by fulvestrant (FIG. 11). In contrast, MCF-7/HER-2 cells wereinsensitive to tamoxifen, but retained partial sensitivity tofulvestrant (28). This correlates with data from the clinic suggestingtamoxifen resistance in breast tumors with high levels of HER-2expression.

Two important systems that transmit extracellular signals into themachinery of the cell nucleus are the signaling pathways that activatenuclear factor κB (NF-κB) and estrogen receptor (ER). These twotranscription factors induce expression of genes that control cellfates, including proliferation and cell death (apoptosis). Estrogenreceptor (ER) and nuclear factor-kappaB (NFκB), a major regulator ofpathways central to malignant progression, are known to be mutuallyinhibitory at several molecular levels. It has been suggested that insome ER-positive breast cancers SERMS such as tamoxifen can activateNFκB, stimulate cell growth and survival, and thereby contribute toendocrine resistance (62). Recent investigations elucidated a previouslyunsuspected effect of ER, namely inhibition of NFκB activation. Inbreast cancer, antiestrogen therapy might cause reactivation of NFκB,potentially re-routing a proliferative signal to breast cancer cells andcontributing to hormone resistance (63). Thus, ligands that selectivelyblock NFκB activation, such as parthenolides, could provide specificpotential therapy for hormone-resistant ER-positive breast cancers (seeFIG. 12). Disclosed herein, inter alia, are solutions to these and otherproblems in the art.

BRIEF SUMMARY OF THE INVENTION

In a first aspect is provided a compound, or a pharmaceuticallyacceptable salt thereof, wherein the compound has a formula selectedfrom the group consisting of:

L⁰ is independently a bond or an unsubstituted C₁-C₁₀ alkylene. R¹ andR² are independently hydrogen, —OH, substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, or substituted or unsubstituted heteroaryl; whereR¹ and R² may optionally be joined to form a substituted orunsubstituted heterocycloalkyl or a substituted or unsubstitutedheteroaryl; with the proviso that the compound, or the pharmaceuticallyacceptable salt thereof is not

In another aspect is provided a pharmaceutical composition including apharmaceutically acceptable excipient and a compound, or apharmaceutically acceptable salt thereof, as described herein (includingembodiments).

In another aspect is provided a method of treating cancer in a patientin need of the treatment, the method including administering atherapeutically effective amount of a compound, or a pharmaceuticallyacceptable salt thereof, as described herein (including embodiments) tothe patient.

In another aspect is provided a method of treating a non-malignanthyperproliferative disease in a patient in need of the treatment, themethod including administering a therapeutically effective amount of acompound, or a pharmaceutically acceptable salt thereof, as describedherein (including embodiments) to the patient.

In another aspect is provided a method of inhibiting cancer cell growthor survival including contacting the cell with an effective amount of acompound, or a pharmaceutically acceptable salt thereof, as describedherein (including embodiments).

In another aspect is provided a method of modulating the level ofactivity of NF-κB in a cell including contacting the cell with aneffective amount of a compound, or a pharmaceutically acceptable saltthereof, as described herein (including embodiments).

In another aspect is provided a method of modulating the level ofactivity of TSC1 in a cell including contacting the cell with aneffective amount of a compound, or a pharmaceutically acceptable saltthereof, as described herein (including embodiments).

In another aspect is provided a method of modulating the level ofactivity of TSC2 in a cell including contacting the cell with aneffective amount of a compound, or a pharmaceutically acceptable saltthereof, as described herein (including embodiments).

In another aspect is provided a method of modulating the level ofactivity of mTOR in a cell including contacting the cell with aneffective amount of a compound, or a pharmaceutically acceptable saltthereof, as described herein (including embodiments).

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Isolation of cancer stem/progenitor cell subsets as determinedby assay of specific biomarkers, ALDH and CD133, and effect of estrogenexposure. NSCLC cells (A549) were treated in vitro with control (CON),10 nM estradiol-17b (E2), or E2 in the presence of 1 mM fulvestrant.Thereafter, aldehyde dehydrogenase (ALDH) activity and CD133 expressionwere analyzed by established double labeling methods using an AldefluorAssay Kit and CD133 antibody, respectively, followed by FACS sorting[11]. The numbers of ALDH+/CD133+ lung tumor cells, representing aputative CSC subset, was expressed as a percent of total lung tumorcells for each treatment group. Therapy with estrogen increases theproportion of cancer stem/progenitor cell subsets as determined by assayof specific biomarkers, ALDH and CD133 (*P<0.001). Experiments were donein triplicate.

FIG. 2. ALDH+/CD133+ tumor cell subpopulations form tumor spheres andexhibit self-renewal properties. For tumor sphere experiments, singlecell suspensions of NSCLC cells (A549) were plated on 1% agarose-coatedplates at a density of 1×10⁵ and grown for 7-10 days. Aldefluor-positive(ALDH+) and CD133-positive (CD133+) cells isolated after treatment withsteroid hormones (31) formed significantly greater numbers and sizes oftumor spheres as compared to Aldefluor-negative (ALDH−) andCD133-negative (CD133−) controls (P<0.001). Subsequent cultures afterdissociation of primary spheres were plated on ultralow attachmentplates at a density of 5×10³ to 1×10⁴ (see P1, P3, 1^(st) and 3^(rd)passages, respectively). Tumor sphere cultures were grown in aserum-free basal medium as previously described [31]. Among ALDH+/CD133+cells isolated after control treatment in the absence of steroidhormones (31), the number of tumor spheres formed was only 11±4 (n=3)per 1000 cells after 7-10 days; this result was significantly differentthan that found with ALDH+/CD133+ cells isolated after treatment withsteroids (P<0.01; not shown). *, values significantly different fromcontrols at P<0.001.

FIG. 3. Parthenolide congener (JD201; DMAPT) elicits dose-dependentincrements in apoptosis of cancer stem cells (CSC) derived from NSCLC(H157). A comparison is performed using bulk NSCLC cells. Results basedon annexin-V assays in 3 independent experiments.

FIG. 4. Parthenolide analogue (JD201; DMAPT) inhibits proliferation ofNSCLC cells using both bulk cell preparations (bulk) andCSC-subpopulations (CSC). Effects are significantly different fromcontrol at P<0.05. JD201 drug dose=20 micromolar.

FIG. 5. Parthenolide analogues JD201-JD210 suppress human lung tumorcell proliferation in vitro. A549, H23, H2122 and H1975 NSCLC cells weretreated with increasing concentrations of drugs (0.1-100 μM). After 72 hcell viability was assessed by MTS assay. Viable cells are expressed aspercentage of vehicle control. P<0.01 (**), versus control group, P<0.05(*), versus control group. Natural occurring parthenolide (PTL),parthenolide analogues JD201 (DMAPT)-JD210. Parthenolide analoguesinhibit female (H1975, H2122) and male (H23, A549) NSCLC cellproliferation. Cells were treated with increasing concentrations ofparental parthenolide (PTL), compounds JD201 (DMAPT/LC-1) and JD203 andcisplatin for comparison.

FIG. 6. Parthenolide analogues sensitize lung tumor cells tocisplatin-induced cytotoxicity. Non-small cell lung cancer cells H23 andA549 were exposed to 10 and 50 μM of each parthenolide analogue compoundin the presence of 5 μM cisplatin. After 72 hrs, cytotoxicity wasmeasured by use of an LDH release assay. Cisplatin cytotoxity at 10 μMis shown as an independent reference control. P<0.01, versus controlgroup. Results represent average values from at least 3 differentexperiments. Parthenolide analogues inhibit female (H1975, H2122) andmale (H23, A549) NSCLC cell proliferation.

FIG. 7. Parthenolide analogues inhibit phosphory-lation of NF-κB/p65.H23 cells were treated with vehicle (CON) or 10 uM concentration ofparthenolide analogues (201, 202, 203, 204, 205). After 60 min Westernblot were performed using anti-phospho-NF-κB/p65 antibody (CellSignaling Technology #3031). Membranes were stripped and re-probed withantibody against total NF-κB/p65 (CST #3037).

FIG. 8. Parthenolide Analogues. Chemical structure of sesquiterpenelactone parthenolide (Parthenolide) and related analogues, DMAPT (JD201)and JD202-210.

FIG. 9. Parthenolide analogues inhibit cisplatin-mediatedphosphorylation of NF-κB/p65; A549 cells were treated with 5 μM ofcisplatin and/or 20 μM of parthenolide analog JD 203 for the varioustime courses displayed. Western blots were then performed usinganti-phospho-NF-κB/p65 antibody (Cell Signaling Technology #3036) andanti-phospho-IκBα antibody (CST #2859). Membranes were then stripped andreprobed with antibody against total NF-κB/p65 (CST #3987) and antibodyagainst total IκBα (CST #4814).

FIG. 10. Mechanisms of estrogen (E2) action; In current models of E2action, E2 binds ER in the nucleus to promote receptor dimerization andphosphorylation that allows binding with coactivator (SRC) andestrogen-responsive elements (ERE), leading to gene transcription (leftpanel). Alternate pathways also impact E2 action. HER-2 receptorsignaling, which may be altered by heregulin (HRG) or other growthfactors, can interact with ER or downstream elements such ascoactivators (middle panel). E2 may also bind ER localized in caveolaeor lipid rafts to promote interaction with HER receptors, G-proteins,nucleotide cyclases, MAP kinase, PI3/Akt kinase or nuclear ER (rightpanel). Reprinted from New Engl J Med, 2002 (40).

FIG. 11. Overexpression of HER2 gene in MCF7 cells elicits resistance toendocrine therapy in vivo; (a) Antiestrogen sensitivity of MCF7 parentcells without HER2 overexpression (control) in vivo (LEFT PANEL). MCF7cells were inoculated SQ in ovari-ectomized, nude mice primed withestrogen. After 10 d, mice were randomized to treatment with tamoxifen(TAM; 5 mg sustained-release pellet SQ), ICI 182,780 (ICI; 5 mg in oilSQ Q wk) or control (CON) for 28d. Tumor volumes were recorded. Nosignificant difference in accumulation of [3H]-tamoxifen by MCF-7 parentvs. HER2 cells was found (n=3; not shown). (b) Antiestrogen sensitivityof HER-2-overexpressing MCF-7 cells (HER-2) in vivo (RIGHT PANEL).MCF-7/HER2 cells were inoculated SQ in ovariectomized, athymic miceprimed with estrogen. After 10 d, animals were randomized to therapywith tamoxifen (TAM), ICI 182,780 (ICI) or control (CON). Refer tomethods as before (28,29).

FIG. 12. Parthenolide analogues that inhibit NFκB activation may play acritical role in reversing therapeutic resistance. Top figure modifiedfrom Kalaitzidis D et al. (36).

FIG. 13. Parthenolide analogues inhibit phosphorylation of NF-κB/p65;H23 cells were treated with vehicle (CON) or 10 mM parthenolide analogs(JD201, 202, 203, 204 and 205); after 60 min, Western blots were doneusing anti-phospho-NFκB/p65 antibody (Cell Signaling); membranes werestripped, re-probed with antibody to total NFκB/p65 (not shown); similarfindings have been obtained using human breast tumor cells in vitro.

FIG. 14. NFkB activity in MCF-7 breast cancer cells is enhanced withtamoxifen resistance and with HER2 overexpression; activity of NFkB inMCF-7 control cells (CON) was compared to that of tamoxifen-resistantMCF-7 (TMR) and MCF-7 HER2-overexpressing (HER2) cells that werecultivated under established in vitro conditions (28,29,67); activity ofNFkB was assessed by using established transient transfection andluciferase reporter assays; cell cultures were seeded one day beforetransfection with luciferase (luc) reporters to a density of 1-2×103cells/well in 96-well plates and using established growth media (67,70);cells were transiently transfected with 0.5 μg of NFκB-luc reportervector (Promega), along with FuGene 6 transfection reagent (Roche);Renilla luciferase vector pRL-tk-luc (Promega) was co-transfected as aninternal control reporter vector to normalize for transfectionefficiency; culture media was changed 20 h following transfection andcells were then maintained in appropriate media for 24 h; as permanufacturer's recommendations, cells were subsequently washed with PBS,lysed for Dual-Glo luciferase assay (Promega), and reporter activitymeasured by luminometer; the ratio of firefly luminescence/Renillaluminescence was used for comparison of NFκB driven gene activities incontrol cells; all transient transfection and reporter gene results arepresented as means (±SEM) from at least 3 independent experiments;MCF7/HER2 and MCF/TMR results showed significant differences (p<0.001)from comparably cultured MCF7 controls.

FIG. 15. Parthenolide analogues 201 and 203 reduce proliferation ofMCF-7/HER2 cells with tamoxifen resistance; analogues 201 and 203 wereboth effective in inducing a dose-dependent (0.1-50 micromolar)reduction in proliferation of MCF-7/HER2 as compared to controls(P<0.001); cell proliferation was determined by cell counts after 72hours incubation in vitro in standard media (28,29,67).

FIG. 16. (A) Parthenolide analogue 203 enhances antitumor efficacy oftamoxifen in tamoxifen-resistant MCF-7/HER2 breast cancer cells;MCF-7/HER2 with tamoxifen resistance were plated in established media(28,67) and then treated with doses of parthenolide analogue 203 rangingfrom 0.1 to 50 micromolar for 72 hours; cell proliferation was thendetermined using cell counts; administration of tamoxifen (1×10-7 M)with parthenolide analogue 203 elicited a greater reduction in cellproliferation than treatment with the analogue alone (P<0.01); (B)Parthenolide analogue 203 promotes antitumor effects of tamoxifen intamoxifen-resistant MCF-7/TAMR breast cancer cells; methods utilized indeveloping tamoxifen-resistant (TAM-R) cells have been described before(64,68); wild-type (wt) and TAM-R MCF-7 cells were plated into 6-wellplates (3×104 cells/well) in RPMI medium containing 5% FBS and grown for2 days; thereafter, use of estrogen-deprived conditions were usedwherein cells were stepped down into phenol red-free media containing 1%dextran-coated charcoal-stripped (DCC) serum (29,67); cells were thentreated with tamoxifen (1×10-7 M) with control vehicle (TM+203 vehicle),analogue 203 alone at varying doses (203 alone) or analogue 203 plustamoxifen (203+TM) for 48 hours; cell numbers were then quantitated asdescribed previously (29,67); experiments were repeated three times toensure reproducibility.

FIG. 17. NFκB activity in SKBR3 breast cancer cells sensitive (CON) andresistant (TZR) to trastuzumab treatment; methods used were as describedin FIG. 14 above.

FIG. 18. Parthenolide analogue 203 reduces proliferation of SKBRS/TZRcells alone and in combination with trastuzumab in vitro; SKBR3 cellswith resistance to trastuzumab were grown with 50 mg/ml trastuzumabalone (TX+203 Vh), parthenolide analogue 203 alone (203) at doses from0.1 to 50 micromolar, or analogue 203 in combination with trastuzumab(203+TZ) for 72 hours prior to cell counts.

FIG. 19. Parthenolide analogue inhibits proliferation of ELT3 andangiomyolipoma (AML) cells that are in vitro models for TSC-relatedproliferative diseases. Cells were counted and plated and then treatedwith parthenolide analogue JD203 or control for 72 hours using IIAmedium with 1% FBS. At the end of the experiments, cell proliferationwas quantitated based on cell counts. Experimental results (n=3) shownas % control (mean±SE).

DETAILED DESCRIPTION I. Definitions

The abbreviations used herein have their conventional meaning within thechemical and biological arts. The chemical structures and formulae setforth herein are constructed according to the standard rules of chemicalvalency known in the chemical arts.

Where substituent groups are specified by their conventional chemicalformulae, written from left to right, they equally encompass thechemically identical substituents that would result from writing thestructure from right to left, e.g., —CH₂O— is equivalent to —OCH₂—.

The term “alkyl,” by itself or as part of another substituent, means,unless otherwise stated, a straight (i.e., unbranched) or branchedcarbon chain (or carbon), or combination thereof, which may be fullysaturated, mono- or polyunsaturated and can include di- and multivalentradicals, having the number of carbon atoms designated (i.e., C₁-C₁₀means one to ten carbons). Examples of saturated hydrocarbon radicalsinclude, but are not limited to, groups such as methyl, ethyl, n-propyl,isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, (cyclohexyl)methyl,homologs and isomers of, for example, n-pentyl, n-hexyl, n-heptyl,n-octyl, and the like. An unsaturated alkyl group is one having one ormore double bonds or triple bonds. Examples of unsaturated alkyl groupsinclude, but are not limited to, vinyl, 2-propenyl, crotyl,2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl),ethynyl, 1- and 3-propynyl, 3-butynyl, and the higher homologs andisomers. An alkoxy is an alkyl attached to the remainder of the moleculevia an oxygen linker (—O—).

The term “alkylene,” by itself or as part of another substituent, means,unless otherwise stated, a divalent radical derived from an alkyl, asexemplified, but not limited by, —CH₂CH₂CH₂CH₂—. Typically, an alkyl (oralkylene) group will have from 1 to 24 carbon atoms, with those groupshaving 10 or fewer carbon atoms being preferred in the presentinvention. A “lower alkyl” or “lower alkylene” is a shorter chain alkylor alkylene group, generally having eight or fewer carbon atoms. Theterm “alkenylene,” by itself or as part of another substituent, means,unless otherwise stated, a divalent radical derived from an alkene.

The term “heteroalkyl,” by itself or in combination with another term,means, unless otherwise stated, a stable straight or branched chain, orcombinations thereof, including at least one carbon atom and at leastone heteroatom selected from the group consisting of O, N, P, Si, and S,and wherein the nitrogen and sulfur atoms may optionally be oxidized,and the nitrogen heteroatom may optionally be quaternized. Theheteroatom(s) O, N, P, S, and Si may be placed at any interior positionof the heteroalkyl group or at the position at which the alkyl group isattached to the remainder of the molecule. Examples include, but are notlimited to: —CH₂—CH₂—O—CH₃, —CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃,—CH₂—S—CH₂—CH₃, —CH₂—CH₂, —S(O)—CH₃, —CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃,—Si(CH₃)₃, —CH₂—CH═N—OCH₃, —CH═CH—N(CH₃)—CH₃, —O—CH₃, —O—CH₂—CH₃, and—CN. Up to two or three heteroatoms may be consecutive, such as, forexample, —CH₂—NH—OCH₃ and —CH₂—O—Si(CH₃)₃.

Similarly, the term “heteroalkylene,” by itself or as part of anothersubstituent, means, unless otherwise stated, a divalent radical derivedfrom heteroalkyl, as exemplified, but not limited by,—CH₂—CH₂—S—CH₂—CH₂— and —CH₂—S—CH₂—CH₂—NH—CH₂—. For heteroalkylenegroups, heteroatoms can also occupy either or both of the chain termini(e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, andthe like). Still further, for alkylene and heteroalkylene linkinggroups, no orientation of the linking group is implied by the directionin which the formula of the linking group is written. For example, theformula —C(O)₂R′— represents both —C(O)₂R′— and —R′C(O)₂—. As describedabove, heteroalkyl groups, as used herein, include those groups that areattached to the remainder of the molecule through a heteroatom, such as—C(O)R′, —C(O)NR′, —NR′R″, —OR′, —SR′, and/or —SO₂R′. Where“heteroalkyl” is recited, followed by recitations of specificheteroalkyl groups, such as —NR′R″ or the like, it will be understoodthat the terms heteroalkyl and —NR′R″ are not redundant or mutuallyexclusive. Rather, the specific heteroalkyl groups are recited to addclarity. Thus, the term “heteroalkyl” should not be interpreted hereinas excluding specific heteroalkyl groups, such as —NR′R″ or the like.

The terms “cycloalkyl” and “heterocycloalkyl,” by themselves or incombination with other terms, mean, unless otherwise stated, cyclicversions of “alkyl” and “heteroalkyl,” respectively, wherein the carbonsmaking up the ring or rings do not necessarily need to be bonded to ahydrogen due to all carbon valencies participating in bonds withnon-hydrogen atoms. Additionally, for heterocycloalkyl, a heteroatom canoccupy the position at which the heterocycle is attached to theremainder of the molecule. Examples of cycloalkyl include, but are notlimited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples ofheterocycloalkyl include, but are not limited to,1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl,3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl,tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl,1-piperazinyl, 2-piperazinyl, and the like. A “cycloalkylene” and a“heterocycloalkylene,” alone or as part of another substituent, means adivalent radical derived from a cycloalkyl and heterocycloalkyl,respectively.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“halo(C₁-C₄)alkyl” includes, but is not limited to, fluoromethyl,difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl,3-bromopropyl, and the like.

The term “acyl” means, unless otherwise stated, —C(O)R where R is asubstituted or unsubstituted alkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl.

The term “aryl” means, unless otherwise stated, a polyunsaturated,aromatic, hydrocarbon substituent, which can be a single ring ormultiple rings (preferably from 1 to 3 rings) that are fused together(i.e., a fused ring aryl) or linked covalently. A fused ring aryl refersto multiple rings fused together wherein at least one of the fused ringsis an aryl ring. The term “heteroaryl” refers to aryl groups (or rings)that contain at least one heteroatom such as N, O, or S, wherein thenitrogen and sulfur atoms are optionally oxidized, and the nitrogenatom(s) are optionally quaternized. Thus, the term “heteroaryl” includesfused ring heteroaryl groups (i.e., multiple rings fused togetherwherein at least one of the fused rings is a heteroaromatic ring). A5,6-fused ring heteroarylene refers to two rings fused together, whereinone ring has 5 members and the other ring has 6 members, and wherein atleast one ring is a heteroaryl ring. Likewise, a 6,6-fused ringheteroarylene refers to two rings fused together, wherein one ring has 6members and the other ring has 6 members, and wherein at least one ringis a heteroaryl ring. And a 6,5-fused ring heteroarylene refers to tworings fused together, wherein one ring has 6 members and the other ringhas 5 members, and wherein at least one ring is a heteroaryl ring. Aheteroaryl group can be attached to the remainder of the moleculethrough a carbon or heteroatom. Non-limiting examples of aryl andheteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl,1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl,4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl,5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl,4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl,2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl,5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl,5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and6-quinolyl. Substituents for each of the above noted aryl and heteroarylring systems are selected from the group of acceptable substituentsdescribed below. An “arylene” and a “heteroarylene,” alone or as part ofanother substituent, mean a divalent radical derived from an aryl andheteroaryl, respectively. Non-limiting examples of heteroaryl groupsinclude pyridinyl, pyrimidinyl, thiophenyl, thienyl, furanyl, indolyl,benzoxadiazolyl, benzodioxolyl, benzodioxanyl, thianaphthanyl,pyrrolopyridinyl, indazolyl, quinolinyl, quinoxalinyl, pyridopyrazinyl,quinazolinonyl, benzoisoxazolyl, imidazopyridinyl, benzofuranyl,benzothienyl, benzothiophenyl, phenyl, naphthyl, biphenyl, pyrrolyl,pyrazolyl, imidazolyl, pyrazinyl, oxazolyl, isoxazolyl, thiazolyl,furylthienyl, pyridyl, pyrimidyl, benzothiazolyl, purinyl,benzimidazolyl, isoquinolyl, thiadiazolyl, oxadiazolyl, pyrrolyl,diazolyl, triazolyl, tetrazolyl, benzothiadiazolyl, isothiazolyl,pyrazolopyrimidinyl, pyrrolopyrimidinyl, benzotriazolyl, benzoxazolyl,or quinolyl. The examples above may be substituted or unsubstituted anddivalent radicals of each heteroaryl example above are non-limitingexamples of heteroarylene.

A fused ring heterocyloalkyl-aryl is an aryl fused to aheterocycloalkyl. A fused ring heterocycloalkyl-heteroaryl is aheteroaryl fused to a heterocycloalkyl. A fused ringheterocycloalkyl-cycloalkyl is a heterocycloalkyl fused to a cycloalkyl.A fused ring heterocycloalkyl-heterocycloalkyl is a heterocycloalkylfused to another heterocycloalkyl. Fused ring heterocycloalkyl-aryl,fused ring heterocycloalkyl-heteroaryl, fused ringheterocycloalkyl-cycloalkyl, or fused ringheterocycloalkyl-heterocycloalkyl may each independently beunsubstituted or substituted with one or more of the substitutentsdescribed herein.

The term “oxo,” as used herein, means an oxygen that is double bonded toa carbon atom.

The term “alkylsulfonyl,” as used herein, means a moiety having theformula —S(O₂)—R′, where R′ is a substituted or unsubstituted alkylgroup as defined above. R′ may have a specified number of carbons (e.g.,“C₁-C₄ alkylsulfonyl”).

Each of the above terms (e.g., “alkyl,” “heteroalkyl,” “aryl,” and“heteroaryl”) includes both substituted and unsubstituted forms of theindicated radical. Preferred substituents for each type of radical areprovided below.

Substituents for the alkyl and heteroalkyl radicals (including thosegroups often referred to as alkylene, alkenyl, heteroalkylene,heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, andheterocycloalkenyl) can be one or more of a variety of groups selectedfrom, but not limited to, —OR′, ═O, ═NR′, ═N—OR′, —NR′R″, —SR′,-halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″,—NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NR—C(NR′R″R′″)═NR″″,—NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NRSO₂R′, —NR′NR″R′″,—ONR′R″, NR′C═(O)NR″NR′″R″″, —CN, —NO₂, monophosphate (or derivativesthereof), diphosphate (or derivatives thereof), triphosphate (orderivatives thereof), in a number ranging from zero to (2m′+1), where m′is the total number of carbon atoms in such radical. R, R′, R″, R′″, andR″″ each preferably independently refer to hydrogen, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl (e.g., aryl substituted with 1-3 halogens),substituted or unsubstituted heteroaryl, substituted or unsubstitutedalkyl, alkoxy, or thioalkoxy groups, or arylalkyl groups. When acompound of the invention includes more than one R group, for example,each of the R groups is independently selected as are each R′, R″, R′″,and R″″ group when more than one of these groups is present. When R′ andR″ are attached to the same nitrogen atom, they can be combined with thenitrogen atom to form a 4-, 5-, 6-, or 7-membered ring. For example,—NR′R″ includes, but is not limited to, 1-pyrrolidinyl and4-morpholinyl. From the above discussion of substituents, one of skillin the art will understand that the term “alkyl” is meant to includegroups including carbon atoms bound to groups other than hydrogengroups, such as haloalkyl (e.g., —CF₃ and —CH₂CF₃) and acyl (e.g.,—C(O)CH₃, —C(O)CF₃, —C(O)CH₂OCH₃, and the like).

Similar to the substituents described for the alkyl radical,substituents for the aryl and heteroaryl groups are varied and areselected from, for example: —OR′, —NR′R″, —SR′, -halogen, —SiR′R″R′″,—OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″, —NR″C(O)R′,—NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″,—S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NRSO₂R′, —NR′NR″R′″, —ONR′R″,—NR′C═(O)NR″NR′″R″″, —CN, —NO₂, —R′, —N₃, —CH(Ph)₂, fluoro(C₁-C₄)alkoxy,and fluoro(C₁-C₄)alkyl, in a number ranging from zero to the totalnumber of open valences on the aromatic ring system; and where R′, R″,R′″, and R″″ are preferably independently selected from hydrogen,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. When a compound of theinvention includes more than one R group, for example, each of the Rgroups is independently selected as are each R′, R″, R′″, and R″″ groupswhen more than one of these groups is present.

Two or more substituents may optionally be joined to form aryl,heteroaryl, cycloalkyl, or heterocycloalkyl groups. Such so-calledring-forming substituents are typically, though not necessarily, foundattached to a cyclic base structure. In one embodiment, the ring-formingsubstituents are attached to adjacent members of the base structure. Forexample, two ring-forming substituents attached to adjacent members of acyclic base structure create a fused ring structure. In anotherembodiment, the ring-forming substituents are attached to a singlemember of the base structure. For example, two ring-forming substituentsattached to a single member of a cyclic base structure create aspirocyclic structure. In yet another embodiment, the ring-formingsubstituents are attached to non-adjacent members of the base structure.

Two of the substituents on adjacent atoms of the aryl or heteroaryl ringmay optionally form a ring of the formula -T-C(O)—(CRR′)_(q)—U—, whereinT and U are independently —NR—, —O—, —CRR′—, or a single bond, and q isan integer of from 0 to 3. Alternatively, two of the substituents onadjacent atoms of the aryl or heteroaryl ring may optionally be replacedwith a substituent of the formula -A-(CH₂)_(r)—B—, wherein A and B areindependently —CRR′—, —O—, —NR—, —S—, —S(O)—, —S(O)₂—, —S(O)₂NR′—, or asingle bond, and r is an integer of from 1 to 4. One of the single bondsof the new ring so formed may optionally be replaced with a double bond.Alternatively, two of the substituents on adjacent atoms of the aryl orheteroaryl ring may optionally be replaced with a substituent of theformula —(CRR′)_(s)—X′—(C′″R″R′″)_(d)—, where s and d are independentlyintegers of from 0 to 3, and X′ is —O—, —NW—, —S—, —S(O)—, —S(O)₂—, or—S(O)₂NR′—. The substituents R, R′, R″, and R′″ are preferablyindependently selected from hydrogen, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, and substituted or unsubstitutedheteroaryl.

As used herein, the terms “heteroatom” or “ring heteroatom” are meant toinclude, oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), andsilicon (Si).

A “substituent group,” as used herein, means a group selected from thefollowing moieties:

-   -   (A) oxo, halogen, —CF₃, —CCl₃, —CN, —OH, —NH₂, —COOH, —CONH₂,        —NO₂, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,        —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH,        —NHOH, —OCF₃, —OCHF₂, unsubstituted alkyl, unsubstituted        heteroalkyl, unsubstituted cycloalkyl, unsubstituted        heterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl,        and    -   (B) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,        heteroaryl, substituted with at least one substituent selected        from:        -   (i) oxo, halogen, —CF₃, —CCl₃, —CN, —OH, —NH₂, —COOH,            —CONH₂, —NO₂, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,            —ONH₂, —NHC═(O)NHNH₂, —NHC═(O) NH₂, —NHSO₂H, —NHC═(O)H,            —NHC(O)—OH, —NHOH, —OCF₃, —OCHF₂, unsubstituted alkyl,            unsubstituted heteroalkyl, unsubstituted cycloalkyl,            unsubstituted heterocycloalkyl, unsubstituted aryl,            unsubstituted heteroaryl, and        -   (ii) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,            heteroaryl, substituted with at least one substituent            selected from:            -   (a) oxo, halogen, —CF₃, —CCl₃, —CN, —OH, —NH₂, —COOH,                —CONH₂, —NO₂, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂,                —NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O) NH₂, —NHSO₂H,                —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCF₃, —OCHF₂,                unsubstituted alkyl, unsubstituted heteroalkyl,                unsubstituted cycloalkyl, unsubstituted                heterocycloalkyl, unsubstituted aryl, unsubstituted                heteroaryl, and            -   (b) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,                aryl, heteroaryl, substituted with at least one                substituent selected from: oxo, halogen, —CF₃, —CCl₃,                —CN, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —SH, —SO₂Cl, —SO₃H,                —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂,                —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH,                —OCF₃, —OCHF₂, unsubstituted alkyl, unsubstituted                heteroalkyl, unsubstituted cycloalkyl, unsubstituted                heterocycloalkyl, unsubstituted aryl, unsubstituted                heteroaryl.

A “size-limited substituent” or “size-limited substituent group,” asused herein, means a group selected from all of the substituentsdescribed above for a “substituent group,” wherein each substituted orunsubstituted alkyl is a substituted or unsubstituted C₁-C₂₀ alkyl, eachsubstituted or unsubstituted heteroalkyl is a substituted orunsubstituted 2 to 20 membered heteroalkyl, each substituted orunsubstituted cycloalkyl is a substituted or unsubstituted C₃-C₈cycloalkyl, each substituted or unsubstituted heterocycloalkyl is asubstituted or unsubstituted 3 to 8 membered heterocycloalkyl, eachsubstituted or unsubstituted aryl is a substituted or unsubstitutedC₆-C₁₀ aryl, and each substituted or unsubstituted heteroaryl is asubstituted or unsubstituted 5 to 10 membered heteroaryl.

A “lower substituent” or “lower substituent group,” as used herein,means a group selected from all of the substituents described above fora “substituent group,” wherein each substituted or unsubstituted alkylis a substituted or unsubstituted C₁-C₈ alkyl, each substituted orunsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8membered heteroalkyl, each substituted or unsubstituted cycloalkyl is asubstituted or unsubstituted C₃-C₇ cycloalkyl, each substituted orunsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 7membered heterocycloalkyl, each substituted or unsubstituted aryl is asubstituted or unsubstituted C₆-C₁₀ aryl, and each substituted orunsubstituted heteroaryl is a substituted or unsubstituted 5 to 9membered heteroaryl.

In some embodiments, each substituted group described in the compoundsherein is substituted with at least one substituent group. Morespecifically, in some embodiments, each substituted alkyl, substitutedheteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl,substituted aryl, substituted heteroaryl, substituted alkylene,substituted heteroalkylene, substituted cycloalkylene, substitutedheterocycloalkylene, substituted arylene, and/or substitutedheteroarylene described in the compounds herein are substituted with atleast one substituent group. In other embodiments, at least one or allof these groups are substituted with at least one size-limitedsubstituent group. In other embodiments, at least one or all of thesegroups are substituted with at least one lower substituent group.

In other embodiments of the compounds herein, each substituted orunsubstituted alkyl may be a substituted or unsubstituted C₁-C₂₀ alkyl,each substituted or unsubstituted heteroalkyl is a substituted orunsubstituted 2 to 20 membered heteroalkyl, each substituted orunsubstituted cycloalkyl is a substituted or unsubstituted C₃-C₈cycloalkyl, each substituted or unsubstituted heterocycloalkyl is asubstituted or unsubstituted 3 to 8 membered heterocycloalkyl, eachsubstituted or unsubstituted aryl is a substituted or unsubstitutedC₆-C₁₀ aryl, and/or each substituted or unsubstituted heteroaryl is asubstituted or unsubstituted 5 to 10 membered heteroaryl. In someembodiments of the compounds herein, each substituted or unsubstitutedalkylene is a substituted or unsubstituted C₁-C₂₀ alkylene, eachsubstituted or unsubstituted heteroalkylene is a substituted orunsubstituted 2 to 20 membered heteroalkylene, each substituted orunsubstituted cycloalkylene is a substituted or unsubstituted C₃-C₈cycloalkylene, each substituted or unsubstituted heterocycloalkylene isa substituted or unsubstituted 3 to 8 membered heterocycloalkylene, eachsubstituted or unsubstituted arylene is a substituted or unsubstitutedC₆-C₁₀ arylene, and/or each substituted or unsubstituted heteroaryleneis a substituted or unsubstituted 5 to 10 membered heteroarylene.

In some embodiments, each substituted or unsubstituted alkyl is asubstituted or unsubstituted C₁-C₈ alkyl, each substituted orunsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8membered heteroalkyl, each substituted or unsubstituted cycloalkyl is asubstituted or unsubstituted C₃-C₇ cycloalkyl, each substituted orunsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 7membered heterocycloalkyl, each substituted or unsubstituted aryl is asubstituted or unsubstituted C₆-C₁₀ aryl, and/or each substituted orunsubstituted heteroaryl is a substituted or unsubstituted 5 to 9membered heteroaryl. In some embodiments, each substituted orunsubstituted alkylene is a substituted or unsubstituted C₁-C₈ alkylene,each substituted or unsubstituted heteroalkylene is a substituted orunsubstituted 2 to 8 membered heteroalkylene, each substituted orunsubstituted cycloalkylene is a substituted or unsubstituted C₃-C₇cycloalkylene, each substituted or unsubstituted heterocycloalkylene isa substituted or unsubstituted 3 to 7 membered heterocycloalkylene, eachsubstituted or unsubstituted arylene is a substituted or unsubstitutedC₆-C₁₀ arylene, and/or each substituted or unsubstituted heteroaryleneis a substituted or unsubstituted 5 to 9 membered heteroarylene. In someembodiments, the compound is a chemical species set forth in theExamples section below.

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds that are prepared with relatively nontoxic acidsor bases, depending on the particular substituents found on thecompounds described herein. When compounds of the present inventioncontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable base additionsalts include sodium, potassium, calcium, ammonium, organic amino, ormagnesium salt, or a similar salt. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, maleic, malonic, benzoic, succinic,suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic,p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Alsoincluded are salts of amino acids such as arginate and the like, andsalts of organic acids like glucuronic or galactunoric acids and thelike (see, e.g., Berge et al., Journal of Pharmaceutical Science 66:1-19(1977)). Certain specific compounds of the present invention containboth basic and acidic functionalities that allow the compounds to beconverted into either base or acid addition salts. Otherpharmaceutically acceptable carriers known to those of skill in the artare suitable for the present invention. Salts tend to be more soluble inaqueous or other protonic solvents that are the corresponding free baseforms. In other cases, the preparation may be a lyophilized powder in 1mM-50 mM histidine, 0.1%-2% sucrose, 2%-7% mannitol at a pH range of 4.5to 5.5, that is combined with buffer prior to use.

Thus, the compounds of the present invention may exist as salts, such aswith pharmaceutically acceptable acids. The present invention includessuch salts. Examples of such salts include hydrochlorides,hydrobromides, sulfates, methanesulfonates, nitrates, maleates,acetates, citrates, fumarates, tartrates (e.g., (+)-tartrates,(−)-tartrates, or mixtures thereof including racemic mixtures),succinates, benzoates, and salts with amino acids such as glutamic acid.These salts may be prepared by methods known to those skilled in theart.

The neutral forms of the compounds are preferably regenerated bycontacting the salt with a base or acid and isolating the parentcompound in the conventional manner. The parent form of the compounddiffers from the various salt forms in certain physical properties, suchas solubility in polar solvents.

In addition to salt forms, the present invention provides compounds,which are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are encompassedwithin the scope of the present invention. Certain compounds of thepresent invention may exist in multiple crystalline or amorphous forms.In general, all physical forms are equivalent for the uses contemplatedby the present invention and are intended to be within the scope of thepresent invention.

As used herein, the term “salt” refers to acid or base salts of thecompounds used in the methods of the present invention. Illustrativeexamples of acceptable salts are mineral acid (hydrochloric acid,hydrobromic acid, phosphoric acid, and the like) salts, organic acid(acetic acid, propionic acid, glutamic acid, citric acid and the like)salts, quaternary ammonium(methyl iodide, ethyl iodide, and the like)salts.

Certain compounds of the present invention possess asymmetric carbonatoms (optical or chiral centers) or double bonds; the enantiomers,racemates, diastereomers, tautomers, geometric isomers, stereoisometricforms that may be defined, in terms of absolute stereochemistry, as (R)-or (S)- or, as (D)- or (L)-for amino acids, and individual isomers areencompassed within the scope of the present invention. The compounds ofthe present invention do not include those which are known in art to betoo unstable to synthesize and/or isolate. The present invention ismeant to include compounds in racemic and optically pure forms.Optically active (R)- and (S)-, or (D)- and (L)-isomers may be preparedusing chiral synthons or chiral reagents, or resolved using conventionaltechniques. When the compounds described herein contain olefinic bondsor other centers of geometric asymmetry, and unless specified otherwise,it is intended that the compounds include both E and Z geometricisomers.

As used herein, the term “isomers” refers to compounds having the samenumber and kind of atoms, and hence the same molecular weight, butdiffering in respect to the structural arrangement or configuration ofthe atoms.

The term “tautomer,” as used herein, refers to one of two or morestructural isomers which exist in equilibrium and which are readilyconverted from one isomeric form to another.

It will be apparent to one skilled in the art that certain compounds ofthis invention may exist in tautomeric forms, all such tautomeric formsof the compounds being within the scope of the invention.

Unless otherwise stated, structures depicted herein are also meant toinclude all stereochemical forms of the structure; i.e., the R and Sconfigurations for each asymmetric center. Therefore, singlestereochemical isomers as well as enantiomeric and diastereomericmixtures of the present compounds are within the scope of the invention.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds which differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement of a hydrogen by a deuterium ortritium, or the replacement of a carbon by ¹³C- or ¹⁴C-enriched carbonare within the scope of this invention.

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine-125 (¹²⁵I), or carbon-14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areencompassed within the scope of the present invention.

The symbol “

” denotes the point of attachment of a chemical moiety to the remainderof a molecule or chemical formula.

The terms “a” or “an,” as used in herein means one or more. In addition,the phrase “substituted with a[n],” as used herein, means the specifiedgroup may be substituted with one or more of any or all of the namedsubstituents. For example, where a group, such as an alkyl or heteroarylgroup, is “substituted with an unsubstituted C₁-C₂₀ alkyl, orunsubstituted 2 to 20 membered heteroalkyl,” the group may contain oneor more unsubstituted C₁-C₂₀ alkyls, and/or one or more unsubstituted 2to 20 membered heteroalkyls. Moreover, where a moiety is substitutedwith an R substituent, the group may be referred to as “R-substituted.”Where a moiety is R-substituted, the moiety is substituted with at leastone R substituent and each R substituent is optionally different.

Descriptions of compounds of the present invention are limited byprinciples of chemical bonding known to those skilled in the art.Accordingly, where a group may be substituted by one or more of a numberof substituents, such substitutions are selected so as to comply withprinciples of chemical bonding and to give compounds which are notinherently unstable and/or would be known to one of ordinary skill inthe art as likely to be unstable under ambient conditions, such asaqueous, neutral, and several known physiological conditions. Forexample, a heterocycloalkyl or heteroaryl is attached to the remainderof the molecule via a ring heteroatom in compliance with principles ofchemical bonding known to those skilled in the art thereby avoidinginherently unstable compounds.

The terms “treating” or “treatment” refers to any indicia of success inthe treatment or amelioration of an injury, disease, pathology orcondition, including any objective or subjective parameter such asabatement; remission; diminishing of symptoms or making the injury,pathology or condition more tolerable to the patient; slowing in therate of degeneration or decline; making the final point of degenerationless debilitating; improving a patient's physical or mental well-being.The treatment or amelioration of symptoms can be based on objective orsubjective parameters; including the results of a physical examination,neuropsychiatric exams, and/or a psychiatric evaluation. For example,certain methods herein treat hyperproliferative disorders, such ascancer (e.g. leukemia, lung cancer, epidermoid carcinoma (i.e.squamous-cell carcinoma), fibrosarcoma, liver cancer (e.g.hepatocellular carcinoma), prostate cancer, kidney cancer (e.g. renalcell carcinoma or urothelial cell carcinoma), lymphoma, breast cancer,urinary bladder cancer, prostate cancer, or therapy resistant cancer) ornon-malignant hyperproliferative diseases (e.g. hamartomatous lesion,angiomyolipoma, lymphangioleiomyomatosis, tuberous sclerosis complex, ahamartia, or a hamartoma). For example certain methods herein treatcancer by decreasing or reducing or preventing the occurrence, growth,metastasis, or progression of cancer or by decreasing or reducing orpreventing a symptom of cancer. Symptoms of cancer (e.g. leukemia, lungcancer, epidermoid carcinoma (i.e. squamous-cell carcinoma),fibrosarcoma, liver cancer (e.g. hepatocellular carcinoma), prostatecancer, kidney cancer (e.g. renal cell carcinoma or urothelial cellcarcinoma), lymphoma, breast cancer, urinary bladder cancer, prostatecancer, or therapy resistant cancer) would be known or may be determinedby a person of ordinary skill in the art. The term “treating” andconjugations thereof, include prevention of an injury, pathology,condition, or disease (e.g. preventing the development of one or moresymptoms of cancer (e.g. leukemia, lung cancer, epidermoid carcinoma(i.e. squamous-cell carcinoma), fibrosarcoma, liver cancer (e.g.hepatocellular carcinoma), prostate cancer, kidney cancer (e.g. renalcell carcinoma or urothelial cell carcinoma), lymphoma, breast cancer,urinary bladder cancer, prostate cancer, or therapy resistant cancer).

An “effective amount” is an amount sufficient to accomplish a statedpurpose (e.g. achieve the effect for which it is administered, treat adisease, reduce enzyme activity, increase enzyme activity, reduce one ormore symptoms of a disease or condition). An example of an “effectiveamount” is an amount sufficient to contribute to the treatment,prevention, or reduction of a symptom or symptoms of a disease, whichcould also be referred to as a “therapeutically effective amount.” A“reduction” of a symptom or symptoms (and grammatical equivalents ofthis phrase) means decreasing of the severity or frequency of thesymptom(s), or elimination of the symptom(s). A “prophylacticallyeffective amount” of a drug is an amount of a drug that, whenadministered to a subject, will have the intended prophylactic effect,e.g., preventing or delaying the onset (or reoccurrence) of an injury,disease, pathology or condition, or reducing the likelihood of the onset(or reoccurrence) of an injury, disease, pathology, or condition, ortheir symptoms. The full prophylactic effect does not necessarily occurby administration of one dose, and may occur only after administrationof a series of doses. Thus, a prophylactically effective amount may beadministered in one or more administrations. An “activity decreasingamount,” as used herein, refers to an amount of antagonist (inhibitor)required to decrease the activity of an enzyme or protein relative tothe absence of the antagonist. An “activity increasing amount,” as usedherein, refers to an amount of agonist (activator) required to increasethe activity of an enzyme or protein relative to the absence of theagonist. A “function disrupting amount,” as used herein, refers to theamount of antagonist (inhibitor) required to disrupt the function of anenzyme or protein relative to the absence of the antagonist. A “functionincreasing amount,” as used herein, refers to the amount of agonist(activator) required to increase the function of an enzyme or proteinrelative to the absence of the agonist. The exact amounts will depend onthe purpose of the treatment, and will be ascertainable by one skilledin the art using known techniques (see, e.g., Lieberman, PharmaceuticalDosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technologyof Pharmaceutical Compounding (1999); Pickar, Dosage Calculations(1999); and Remington: The Science and Practice of Pharmacy, 20thEdition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).

The term “associated” or “associated with” in the context of a substanceor substance activity or function associated with a disease (e.g. cancer(e.g. leukemia, lung cancer, epidermoid carcinoma (i.e. squamous-cellcarcinoma), fibrosarcoma, liver cancer (e.g. hepatocellular carcinoma),prostate cancer, kidney cancer (e.g. renal cell carcinoma or urothelialcell carcinoma), lymphoma, breast cancer, urinary bladder cancer,prostate cancer, or therapy resistant cancer) means that the disease(e.g. cancer (e.g. leukemia, lung cancer, epidermoid carcinoma (i.e.squamous-cell carcinoma), fibrosarcoma, liver cancer (e.g.hepatocellular carcinoma), prostate cancer, kidney cancer (e.g. renalcell carcinoma or urothelial cell carcinoma), lymphoma, breast cancer,urinary bladder cancer, prostate cancer, or therapy resistant cancer))is caused by (in whole or in part), or a symptom of the disease iscaused by (in whole or in part) the substance or substance activity orfunction. For example, a symptom of a disease or condition associatedwith an increase in NFκB activity may be a symptom that results(entirely or partially) from an increase in NFκB activity (e.g. increasein NFκB phosphorylation or activity of phosphorylated NFκB or activityof NFκB or increase in activity of an NFκB signal transduction orsignaling pathway). As used herein, what is described as beingassociated with a disease, if a causative agent, could be a target fortreatment of the disease. For example, a disease associated withincreased NFκB activity or NFκB pathway activity (e.g. phosphorylatedNFκB activity or pathway), may be treated with an agent (e.g. compoundas described herein) effective for decreasing the level of activity ofNFκB activity or NFκB pathway or phosphorylated NFκB activity orpathway. For example, a disease associated with phosphorylated NFκB, maybe treated with an agent (e.g. compound as described herein) effectivefor decreasing the level of activity of phosphorylated NFκB or adownstream component or effector of phosphorylated NFκB. For example, adisease associated with NFκB, may be treated with an agent (e.g.compound as described herein) effective for decreasing the level ofactivity of NFκB or a downstream component or effector of NFκB.

“Control” or “control experiment” is used in accordance with its plainordinary meaning and refers to an experiment in which the subjects orreagents of the experiment are treated as in a parallel experimentexcept for omission of a procedure, reagent, or variable of theexperiment. In some instances, the control is used as a standard ofcomparison in evaluating experimental effects.

“Contacting” is used in accordance with its plain ordinary meaning andrefers to the process of allowing at least two distinct species (e.g.chemical compounds including biomolecules, or cells) to becomesufficiently proximal to react, interact or physically touch. It shouldbe appreciated, however, that the resulting reaction product can beproduced directly from a reaction between the added reagents or from anintermediate from one or more of the added reagents which can beproduced in the reaction mixture. The term “contacting” may includeallowing two species to react, interact, or physically touch, whereinthe two species may be a compound as described herein and a protein orenzyme (e.g. NFκB, phosphorylated NFκB, mTOR, mTORC1, TSC1, TSC2 orcomponent of NFκB pathway, phosphorylated NFκB pathway, pathwayactivated by NFκB phosphorylation, mTOR pathway, mTORC1 pathway, TSC1pathway, or TSC2 pathway). In some embodiments contacting includesallowing a compound described herein to interact with a protein orenzyme that is involved in a signaling pathway (e.g. NFκB pathway,phosphorylated NFκB pathway, pathway activated by NFκB phosphorylation,mTOR pathway, mTORC1 pathway, TSC1 pathway, or TSC2 pathway).

As defined herein, the term “inhibition”, “inhibit”, “inhibiting” andthe like in reference to a protein-inhibitor (e.g. antagonist)interaction means negatively affecting (e.g. decreasing) the activity orfunction of the protein relative to the activity or function of theprotein in the absence of the inhibitor. In some embodiments inhibitionrefers to reduction of a disease or symptoms of disease. In someembodiments, inhibition refers to a reduction in the activity of asignal transduction pathway or signaling pathway. Thus, inhibitionincludes, at least in part, partially or totally blocking stimulation,decreasing, preventing, or delaying activation, or inactivating,desensitizing, or down-regulating signal transduction or enzymaticactivity or the amount of a protein. In some embodiments, inhibitionrefers to a decrease in the activity of a signal transduction pathway orsignaling pathway (e.g. NFκB or phosphorylated NFκB or NFκB pathway orphosphorylated NFκB pathway or pathway activated by NFκB phosphorylationor mTOR pathway or mTOR or mTORC1 pathway or mTORC1 or TSC1 or TSC1pathway or TSC2 or TSC2 pathway). Thus, inhibition may include, at leastin part, partially or totally decreasing stimulation, decreasing orreducing activation, or inactivating, desensitizing, or down-regulatingsignal transduction or enzymatic activity or the amount of a proteinincreased in a disease (e.g. level of NFκB activity or protein or levelof phosphorylated NFκB or level or activity of a component of an NFκBpathway or level of phosphorylated NFκB activity or protein or level oractivity of a component of a phosphorylated NFκB pathway or mTOR pathwayor mTOR or mTORC1 pathway or mTORC1 or TSC1 or TSC1 pathway or TSC2 orTSC2 pathway, wherein each is associated with a hyperproliferativedisease, for example cancer (e.g. leukemia, lung cancer, epidermoidcarcinoma (i.e. squamous-cell carcinoma), fibrosarcoma, liver cancer(e.g. hepatocellular carcinoma), prostate cancer, kidney cancer (e.g.renal cell carcinoma or urothelial cell carcinoma), lymphoma, breastcancer, urinary bladder cancer, prostate cancer, or therapy resistantcancer) or a non-malignant hyperproliferative disease (e.g.hamartomatous lesion, angiomyolipoma, lymphangioleiomyomatosis, tuberoussclerosis complex, a hamartia, or a hamartoma). Inhibition may include,at least in part, partially or totally decreasing stimulation,decreasing or reducing activation, or deactivating, desensitizing, ordown-regulating signal transduction or enzymatic activity or the amountof a protein (e.g. NFκB or phosphorylated NFκB or NFκB pathway orphosphorylated NFκB pathway or pathway activated by NFκB phosphorylationor mTOR pathway or mTOR or mTORC1 pathway or mTORC1 or TSC1 or TSC1pathway or TSC2 or TSC2 pathway) that may modulate the level of anotherprotein or increase cell survival (e.g. decrease in phosphorylated NFκBpathway activity may increase cell survival in cells that may or may nothave an increase in phosphorylated NFκB pathway activity relative to anon-disease control or decrease in NFκB pathway activity may increasecell survival in cells that may or may not have an increase in NFκBpathway activity relative to a non-disease control).

As defined herein, the term “activation”, “activate”, “activating” andthe like in reference to a protein-activator (e.g. agonist) interactionmeans positively affecting (e.g. increasing) the activity or function ofthe protein (e.g. NFκB or phosphorylated NFκB or NFκB pathway orphosphorylated NFκB pathway or pathway activated by NFκB phosphorylationor mTOR pathway or mTOR or mTORC1 pathway or mTORC1 or TSC1 or TSC1pathway or TSC2 or TSC2 pathway) relative to the activity or function ofthe protein in the absence of the activator (e.g. compound describedherein). In some embodiments, activation refers to an increase in theactivity of a signal transduction pathway or signaling pathway (e.g.NFκB pathway or phosphorylated NFκB pathway or pathway activated by NFκBphosphorylation or mTOR pathway or mTORC1 pathway or TSC1 pathway orTSC2 pathway). Thus, activation may include, at least in part, partiallyor totally increasing stimulation, increasing or enabling activation, oractivating, sensitizing, or up-regulating signal transduction orenzymatic activity or the amount of a protein decreased in a disease(e.g. level of NFκB activity or level of protein or activity decreasedby phosphorylation of NFκB or protein associated with cancer (e.g.leukemia, lung cancer, epidermoid carcinoma (i.e. squamous-cellcarcinoma), fibrosarcoma, liver cancer (e.g. hepatocellular carcinoma),prostate cancer, kidney cancer (e.g. renal cell carcinoma or urothelialcell carcinoma), lymphoma, breast cancer, urinary bladder cancer,prostate cancer, or therapy resistant cancer). Activation may include,at least in part, partially or totally increasing stimulation,increasing or enabling activation, or activating, sensitizing, orup-regulating signal transduction or enzymatic activity or the amount ofa protein (e.g. NFκB or phosphorylated NFκB or NFκB pathway orphosphorylated NFκB pathway or pathway activated by NFκB phosphorylationor mTOR pathway or mTOR or mTORC1 pathway or mTORC1 or TSC1 or TSC1pathway or TSC2 or TSC2 pathway) that may modulate the level of anotherprotein or increase cell survival (e.g. increase in NFκB activity mayincrease cell survival in cells that may or may not have a reduction inNFκB activity relative to a non-disease control).

The term “modulator” refers to a composition that increases or decreasesthe level of a target molecule or the function of a target molecule. Insome embodiments, a modulator of NFκB or phosphorylated NFκB or NFκBpathway or phosphorylated NFκB pathway or pathway activated by NFκBphosphorylation or mTOR pathway or mTOR or mTORC1 pathway or mTORC1 orTSC1 or TSC1 pathway or TSC2 or TSC2 pathway is a compound that reducesthe severity of one or more symptoms of a disease associated with NFκBor phosphorylated NFκB or NFκB pathway or phosphorylated NFκB pathway orpathway activated by NFκB phosphorylation or mTOR pathway or mTOR ormTORC1 pathway or mTORC1 or TSC1 or TSC1 pathway or TSC2 or TSC2 pathway(e.g. disease associated with an increase or decrease in the level ofNFκB or phosphorylated NFκB or NFκB pathway or phosphorylated NFκBpathway or pathway activated by NFκB phosphorylation or mTOR pathway ormTOR or mTORC1 pathway or mTORC1 or TSC1 or TSC1 pathway or TSC2 or TSC2pathway, for example cancer (e.g. leukemia, lung cancer, epidermoidcarcinoma (i.e. squamous-cell carcinoma), fibrosarcoma, liver cancer(e.g. hepatocellular carcinoma), prostate cancer, kidney cancer (e.g.renal cell carcinoma or urothelial cell carcinoma), lymphoma, breastcancer, urinary bladder cancer, prostate cancer, or therapy resistantcancer) or a non-malignant hyperproliferative disease (e.g.hamartomatous lesion, angiomyolipoma, lymphangioleiomyomatosis, tuberoussclerosis complex, a hamartia, or a hamartoma) or a disease that is notcaused by NFκB or phosphorylated NFκB or NFκB pathway or phosphorylatedNFκB pathway or pathway activated by NFκB phosphorylation or mTORpathway or mTOR or mTORC1 pathway or mTORC1 or TSC1 or TSC1 pathway orTSC2 or TSC2 pathway but may benefit from modulation of NFκB orphosphorylated NFκB or NFκB pathway or phosphorylated NFκB pathway orpathway activated by NFκB phosphorylation or mTOR pathway or mTOR ormTORC1 pathway or mTORC1 or TSC1 or TSC1 pathway or TSC2 or TSC2 pathwayactivity (e.g. decreasing or increasing in level or level of activity ofNFκB or phosphorylated NFκB or NFκB pathway or phosphorylated NFκBpathway or pathway activated by NFκB phosphorylation or mTOR pathway ormTOR or mTORC1 pathway or mTORC1 or TSC1 or TSC1 pathway or TSC2 or TSC2pathway). In embodiments, a modulator of NFκB or phosphorylated NFκB orNFκB pathway or phosphorylated NFκB pathway or pathway activated by NFκBphosphorylation or mTOR pathway or mTOR or mTORC1 pathway or mTORC1 orTSC1 or TSC1 pathway or TSC2 or TSC2 pathway is an anti-cancer agent.

“Patient” or “subject in need thereof” refers to a living organismsuffering from or prone to a disease or condition that can be treated byadministration of a compound or pharmaceutical composition, as providedherein. Non-limiting examples include humans, other mammals, bovines,rats, mice, dogs, monkeys, goat, sheep, cows, deer, and othernon-mammalian animals. In some embodiments, a patient is human.

“Disease” or “condition” refer to a state of being or health status of apatient or subject capable of being treated with a compound,pharmaceutical composition, or method provided herein. In someembodiments, the disease is a disease related to (e.g. caused by) anincrease in the level of activity or amount of NFκB or phosphorylatedNFκB or NFκB pathway or phosphorylated NFκB pathway or pathway activatedby NFκB phosphorylation or mTOR pathway or mTOR or mTORC1 pathway ormTORC1 or TSC1 or TSC1 pathway or TSC2 or TSC2 pathway. In someembodiments, the disease is a disease related to (e.g. caused by) adecrease in the level of activity or amount of NFκB or phosphorylatedNFκB or NFκB pathway or phosphorylated NFκB pathway or pathway activatedby NFκB phosphorylation or mTOR pathway or mTOR or mTORC1 pathway ormTORC1 or TSC1 or TSC1 pathway or TSC2 or TSC2 pathway. In someembodiments, the disease is cancer (e.g. leukemia, lung cancer,epidermoid carcinoma (i.e. squamous-cell carcinoma), fibrosarcoma, livercancer (e.g. hepatocellular carcinoma), prostate cancer, kidney cancer(e.g. renal cell carcinoma or urothelial cell carcinoma), lymphoma,breast cancer, urinary bladder cancer, prostate cancer, or therapyresistant cancer). In some embodiments, the disease is a non-malignanthyperproliferative disease (e.g. hamartomatous lesion, angiomyolipoma,lymphangioleiomyomatosis, tuberous sclerosis complex, a hamartia, or ahamartoma),

Examples of diseases, disorders, or conditions include, but are notlimited to, cancer (e.g. leukemia, lung cancer, epidermoid carcinoma(i.e. squamous-cell carcinoma), fibrosarcoma, liver cancer (e.g.hepatocellular carcinoma), prostate cancer, kidney cancer (e.g. renalcell carcinoma or urothelial cell carcinoma), lymphoma, breast cancer,urinary bladder cancer, prostate cancer, or therapy resistant cancer).In some instances, “disease” or “condition” refers to cancer. In somefurther instances, “cancer” refers to human cancers and carcinomas,sarcomas, adenocarcinomas, lymphomas, leukemias, melanomas, etc.,including solid and lymphoid cancers, kidney, breast, lung, bladder,colon, ovarian, prostate, pancreas, stomach, brain, head and neck, skin,uterine, testicular, glioma, esophagus, liver cancer, includinghepatocarcinoma, lymphoma, including B-acute lymphoblastic lymphoma,non-Hodgkin's lymphomas (e.g., Burkitt's, Small Cell, and Large Celllymphomas), Hodgkin's lymphoma, leukemia (including AML, ALL, and CML),and/or multiple myeloma. In some further instances, “cancer” refers tolung cancer, breast cancer, ovarian cancer, leukemia, lymphoma,melanoma, pancreatic cancer, sarcoma, bladder cancer, bone cancer, braincancer, cervical cancer, colon cancer, esophageal cancer, gastriccancer, liver cancer, head and neck cancer, kidney cancer, myeloma,thyroid cancer, prostate cancer, metastatic cancer, or carcinoma. Inembodiments “cancer” refers to a cancer resistant to an anti-cancertherapy (e.g. treatment with an anti-cancer agent (e.g. platinum-basedcompound, cisplatin, carboplatin, hormonal therapy, hormonal therapeuticagent, tamoxifen, trastuzumab, or an aromatase inhibitor)).

As used herein, the term “cancer” refers to all types of cancer,neoplasm or malignant tumors found in mammals, including leukemia,lymphoma, carcinomas and sarcomas. Exemplary cancers that may be treatedwith a compound, pharmaceutical composition, or method provided hereininclude lymphoma, sarcoma, bladder cancer, bone cancer, brain tumor,cervical cancer, colon cancer, esophageal cancer, gastric cancer, headand neck cancer, kidney cancer, myeloma, thyroid cancer, leukemia,prostate cancer, breast cancer (e.g. ER positive, ER negative,chemotherapy resistant, herceptin resistant, HER2 positive, doxorubicinresistant, tamoxifen resistant, ductal carcinoma, lobular carcinoma,primary, metastatic), ovarian cancer, pancreatic cancer, liver cancer(e.g. hepatocellular carcinoma), lung cancer (e.g. non-small cell lungcarcinoma, squamous cell lung carcinoma, adenocarcinoma, large cell lungcarcinoma, small cell lung carcinoma, carcinoid, sarcoma, cisplatinresistant lung cancer, carboplatin resistant lung cancer, platinum-basedcompound resistant lung cancer), glioblastoma multiforme, glioma, ormelanoma. Additional examples include, cancer of the thyroid, endocrinesystem, brain, breast, cervix, colon, head & neck, liver, kidney, lung,non-small cell lung, melanoma, mesothelioma, ovary, sarcoma, stomach,uterus or Medulloblastoma, Hodgkin's Disease, Non-Hodgkin's Lymphoma,multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme,ovarian cancer, rhabdomyosarcoma, primary thrombocytosis, primarymacroglobulinemia, primary brain tumors, cancer, malignant pancreaticinsulanoma, malignant carcinoid, urinary bladder cancer, premalignantskin lesions, testicular cancer, lymphomas, thyroid cancer,neuroblastoma, esophageal cancer, genitourinary tract cancer, malignanthypercalcemia, endometrial cancer, adrenal cortical cancer, neoplasms ofthe endocrine or exocrine pancreas, medullary thyroid cancer, medullarythyroid carcinoma, melanoma, colorectal cancer, papillary thyroidcancer, hepatocellular carcinoma, Paget's Disease of the Nipple,Phyllodes Tumors, Lobular Carcinoma, Ductal Carcinoma, cancer of thepancreatic stellate cells, cancer of the hepatic stellate cells, orprostate cancer.

The term “leukemia” refers broadly to progressive, malignant diseases ofthe blood-forming organs and is generally characterized by a distortedproliferation and development of leukocytes and their precursors in theblood and bone marrow. Leukemia is generally clinically classified onthe basis of (1) the duration and character of the disease-acute orchronic; (2) the type of cell involved; myeloid (myelogenous), lymphoid(lymphogenous), or monocytic; and (3) the increase or non-increase inthe number abnormal cells in the blood-leukemic or aleukemic(subleukemic). Exemplary leukemias that may be treated with a compound,pharmaceutical composition, or method provided herein include, forexample, acute nonlymphocytic leukemia, chronic lymphocytic leukemia,acute granulocytic leukemia, chronic granulocytic leukemia, acutepromyelocytic leukemia, adult T-cell leukemia, aleukemic leukemia, aleukocythemic leukemia, basophylic leukemia, blast cell leukemia, bovineleukemia, chronic myelocytic leukemia, leukemia cutis, embryonalleukemia, eosinophilic leukemia, Gross' leukemia, hairy-cell leukemia,hemoblastic leukemia, hemocytoblastic leukemia, histiocytic leukemia,stem cell leukemia, acute monocytic leukemia, leukopenic leukemia,lymphatic leukemia, lymphoblastic leukemia, lymphocytic leukemia,lymphogenous leukemia, lymphoid leukemia, lymphosarcoma cell leukemia,mast cell leukemia, megakaryocytic leukemia, micromyeloblastic leukemia,monocytic leukemia, myeloblastic leukemia, myelocytic leukemia, myeloidgranulocytic leukemia, myelomonocytic leukemia, Naegeli leukemia, plasmacell leukemia, multiple myeloma, plasmacytic leukemia, promyelocyticleukemia, Rieder cell leukemia, Schilling's leukemia, stem cellleukemia, subleukemic leukemia, or undifferentiated cell leukemia.

The term “sarcoma” generally refers to a tumor which is made up of asubstance like the embryonic connective tissue and is generally composedof closely packed cells embedded in a fibrillar or homogeneoussubstance. Sarcomas that may be treated with a compound, pharmaceuticalcomposition, or method provided herein include a chondrosarcoma,fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma,Abemethy's sarcoma, adipose sarcoma, liposarcoma, alveolar soft partsarcoma, ameloblastic sarcoma, botryoid sarcoma, chloroma sarcoma,chorio carcinoma, embryonal sarcoma, Wilms' tumor sarcoma, endometrialsarcoma, stromal sarcoma, Ewing's sarcoma, fascial sarcoma, fibroblasticsarcoma, giant cell sarcoma, granulocytic sarcoma, Hodgkin's sarcoma,idiopathic multiple pigmented hemorrhagic sarcoma, immunoblastic sarcomaof B cells, lymphoma, immunoblastic sarcoma of T-cells, Jensen'ssarcoma, Kaposi's sarcoma, Kupffer cell sarcoma, angiosarcoma,leukosarcoma, malignant mesenchymoma sarcoma, parosteal sarcoma,reticulocytic sarcoma, Rous sarcoma, serocystic sarcoma, synovialsarcoma, or telangiectaltic sarcoma.

The term “melanoma” is taken to mean a tumor arising from themelanocytic system of the skin and other organs. Melanomas that may betreated with a compound, pharmaceutical composition, or method providedherein include, for example, acral-lentiginous melanoma, amelanoticmelanoma, benign juvenile melanoma, Cloudman's melanoma, S91 melanoma,Harding-Passey melanoma, juvenile melanoma, lentigo maligna melanoma,malignant melanoma, nodular melanoma, subungal melanoma, or superficialspreading melanoma.

The term “carcinoma” refers to a malignant new growth made up ofepithelial cells tending to infiltrate the surrounding tissues and giverise to metastases. Exemplary carcinomas that may be treated with acompound, pharmaceutical composition, or method provided herein include,for example, medullary thyroid carcinoma, familial medullary thyroidcarcinoma, acinar carcinoma, acinous carcinoma, adenocystic carcinoma,adenoid cystic carcinoma, carcinoma adenomatosum, carcinoma of adrenalcortex, alveolar carcinoma, alveolar cell carcinoma, basal cellcarcinoma, carcinoma basocellulare, basaloid carcinoma, basosquamouscell carcinoma, bronchioalveolar carcinoma, bronchiolar carcinoma,bronchogenic carcinoma, cerebriform carcinoma, cholangiocellularcarcinoma, chorionic carcinoma, colloid carcinoma, comedo carcinoma,corpus carcinoma, cribriform carcinoma, carcinoma en cuirasse, carcinomacutaneum, cylindrical carcinoma, cylindrical cell carcinoma, ductcarcinoma, ductal carcinoma, carcinoma durum, embryonal carcinoma,encephaloid carcinoma, epiermoid carcinoma, carcinoma epithelialeadenoides, exophytic carcinoma, carcinoma ex ulcere, carcinoma fibrosum,gelatiniforni carcinoma, gelatinous carcinoma, giant cell carcinoma,carcinoma gigantocellulare, glandular carcinoma, granulosa cellcarcinoma, hair-matrix carcinoma, hematoid carcinoma, hepatocellularcarcinoma, Hurthle cell carcinoma, hyaline carcinoma, hypernephroidcarcinoma, infantile embryonal carcinoma, carcinoma in situ,intraepidermal carcinoma, intraepithelial carcinoma, Krompecher'scarcinoma, Kulchitzky-cell carcinoma, large-cell carcinoma, lenticularcarcinoma, carcinoma lenticulare, lipomatous carcinoma, lobularcarcinoma, lymphoepithelial carcinoma, carcinoma medullare, medullarycarcinoma, melanotic carcinoma, carcinoma molle, mucinous carcinoma,carcinoma muciparum, carcinoma mucocellulare, mucoepidermoid carcinoma,carcinoma mucosum, mucous carcinoma, carcinoma myxomatodes,nasopharyngeal carcinoma, oat cell carcinoma, carcinoma ossificans,osteoid carcinoma, papillary carcinoma, periportal carcinoma,preinvasive carcinoma, prickle cell carcinoma, pultaceous carcinoma,renal cell carcinoma of kidney, reserve cell carcinoma, carcinomasarcomatodes, schneiderian carcinoma, scirrhous carcinoma, carcinomascroti, signet-ring cell carcinoma, carcinoma simplex, small-cellcarcinoma, solanoid carcinoma, spheroidal cell carcinoma, spindle cellcarcinoma, carcinoma spongiosum, squamous carcinoma, squamous cellcarcinoma, string carcinoma, carcinoma telangiectaticum, carcinomatelangiectodes, transitional cell carcinoma, carcinoma tuberosum,tubular carcinoma, tuberous carcinoma, verrucous carcinoma, or carcinomavillosum.

The term “non-malignant hyperproliferative disease” is used inaccordance with its plain ordinary meaning and refers to a diseaseincluding a growth that is not cancerous. Examples of a non-malignanthyperproliferative disease include a hamartomatous lesion,angiomyolipoma, lymphangioleiomyomatosis, tuberous sclerosis complex, ahamartia, or a hamartoma.

The term “signaling pathway” as used herein refers to a series ofinteractions between cellular and optionally extra-cellular components(e.g. proteins, nucleic acids, small molecules, ions, lipids) thatconveys a change in one component to one or more other components, whichin turn may convey a change to additional components, which isoptionally propagated to other signaling pathway components.

“Pharmaceutically acceptable excipient” and “pharmaceutically acceptablecarrier” refer to a substance that aids the administration of an activeagent to and absorption by a subject and can be included in thecompositions of the present invention without causing a significantadverse toxicological effect on the patient. Non-limiting examples ofpharmaceutically acceptable excipients include water, NaCl, normalsaline solutions, lactated Ringer's, normal sucrose, normal glucose,binders, fillers, disintegrants, lubricants, coatings, sweeteners,flavors, salt solutions (such as Ringer's solution), alcohols, oils,gelatins, carbohydrates such as lactose, amylose or starch, fatty acidesters, hydroxymethycellulose, polyvinyl pyrrolidine, and colors, andthe like. Such preparations can be sterilized and, if desired, mixedwith auxiliary agents such as lubricants, preservatives, stabilizers,wetting agents, emulsifiers, salts for influencing osmotic pressure,buffers, coloring, and/or aromatic substances and the like that do notdeleteriously react with the compounds of the invention. One of skill inthe art will recognize that other pharmaceutical excipients are usefulin the present invention.

The term “preparation” is intended to include the formulation of theactive compound with encapsulating material as a carrier providing acapsule in which the active component with or without other carriers, issurrounded by a carrier, which is thus in association with it.Similarly, cachets and lozenges are included. Tablets, powders,capsules, pills, cachets, and lozenges can be used as solid dosage formssuitable for oral administration.

As used herein, the term “administering” means oral administration,administration as a suppository, topical contact, intravenous,parenteral, intraperitoneal, intramuscular, intralesional, intrathecal,intracranial, intranasal or subcutaneous administration, or theimplantation of a slow-release device, e.g., a mini-osmotic pump, to asubject. Administration is by any route, including parenteral andtransmucosal (e.g., buccal, sublingual, palatal, gingival, nasal,vaginal, rectal, or transdermal). Parenteral administration includes,e.g., intravenous, intramuscular, intra-arteriole, intradermal,subcutaneous, intraperitoneal, intraventricular, and intracranial. Othermodes of delivery include, but are not limited to, the use of liposomalformulations, intravenous infusion, transdermal patches, etc. By“co-administer” it is meant that a composition described herein isadministered at the same time, just prior to, or just after theadministration of one or more additional therapies (e.g. anti-canceragent or chemotherapeutic). The compound of the invention can beadministered alone or can be co-administered to the patient.Co-administration is meant to include simultaneous or sequentialadministration of the compound individually or in combination (more thanone compound or agent). Thus, the preparations can also be combined,when desired, with other active substances (e.g. to reduce metabolicdegradation). The compositions of the present invention can be deliveredby transdermally, by a topical route, formulated as applicator sticks,solutions, suspensions, emulsions, gels, creams, ointments, pastes,jellies, paints, powders, and aerosols. Oral preparations includetablets, pills, powder, dragees, capsules, liquids, lozenges, cachets,gels, syrups, slurries, suspensions, etc., suitable for ingestion by thepatient. Solid form preparations include powders, tablets, pills,capsules, cachets, suppositories, and dispersible granules. Liquid formpreparations include solutions, suspensions, and emulsions, for example,water or water/propylene glycol solutions. The compositions of thepresent invention may additionally include components to providesustained release and/or comfort. Such components include high molecularweight, anionic mucomimetic polymers, gelling polysaccharides andfinely-divided drug carrier substrates. These components are discussedin greater detail in U.S. Pat. Nos. 4,911,920; 5,403,841; 5,212,162; and4,861,760. The entire contents of these patents are incorporated hereinby reference in their entirety for all purposes. The compositions of thepresent invention can also be delivered as microspheres for slow releasein the body. For example, microspheres can be administered viaintradermal injection of drug-containing microspheres, which slowlyrelease subcutaneously (see Rao, J. Biomater Sci. Polym. Ed. 7:623-645,1995; as biodegradable and injectable gel formulations (see, e.g., GaoPharm. Res. 12:857-863, 1995); or, as microspheres for oraladministration (see, e.g., Eyles, J. Pharm. Pharmacol. 49:669-674,1997). In another embodiment, the formulations of the compositions ofthe present invention can be delivered by the use of liposomes whichfuse with the cellular membrane or are endocytosed, i.e., by employingreceptor ligands attached to the liposome, that bind to surface membraneprotein receptors of the cell resulting in endocytosis. By usingliposomes, particularly where the liposome surface carries receptorligands specific for target cells, or are otherwise preferentiallydirected to a specific organ, one can focus the delivery of thecompositions of the present invention into the target cells in vivo.(See, e.g., Al-Muhammed, J. Microencapsul. 13:293-306, 1996; Chonn,Curr. Opin. Biotechnol. 6:698-708, 1995; Ostro, Am. J. Hosp. Pharm.46:1576-1587, 1989). The compositions of the present invention can alsobe delivered as nanoparticles.

Pharmaceutical compositions provided by the present invention includecompositions wherein the active ingredient (e.g. compounds describedherein, including embodiments or examples) is contained in atherapeutically effective amount, i.e., in an amount effective toachieve its intended purpose. The actual amount effective for aparticular application will depend, inter alia, on the condition beingtreated. When administered in methods to treat a disease, suchcompositions will contain an amount of active ingredient effective toachieve the desired result, e.g., modulating the activity of a targetmolecule (e.g. NFκB, phosphorylated NFκB, mTOR, mTORC1, TSC1, TSC2 orcomponent of NFκB pathway, phosphorylated NFκB pathway, pathwayactivated by NFκB phosphorylation, mTOR pathway, mTORC1 pathway, TSC1pathway, or TSC2 pathway), and/or reducing, eliminating, or slowing theprogression of disease symptoms (e.g. symptoms of cancer (e.g. leukemia,lung cancer, epidermoid carcinoma (i.e. squamous-cell carcinoma),fibrosarcoma, liver cancer (e.g. hepatocellular carcinoma), prostatecancer, kidney cancer (e.g. renal cell carcinoma or urothelial cellcarcinoma), lymphoma, breast cancer, urinary bladder cancer, prostatecancer, or therapy resistant cancer) or a non-malignanthyperproliferative disease (e.g. hamartomatous lesion, angiomyolipoma,lymphangioleiomyomatosis, tuberous sclerosis complex, a hamartia, or ahamartoma)). Determination of a therapeutically effective amount of acompound of the invention is well within the capabilities of thoseskilled in the art, especially in light of the detailed disclosureherein.

The dosage and frequency (single or multiple doses) administered to amammal can vary depending upon a variety of factors, for example,whether the mammal suffers from another disease, and its route ofadministration; size, age, sex, health, body weight, body mass index,and diet of the recipient; nature and extent of symptoms of the diseasebeing treated (e.g. symptoms of cancer (e.g. leukemia, lung cancer,epidermoid carcinoma (i.e. squamous-cell carcinoma), fibrosarcoma, livercancer (e.g. hepatocellular carcinoma), prostate cancer, kidney cancer(e.g. renal cell carcinoma or urothelial cell carcinoma), lymphoma,breast cancer, urinary bladder cancer, prostate cancer, or therapyresistant cancer) or a non-malignant hyperproliferative disease (e.g.hamartomatous lesion, angiomyolipoma, lymphangioleiomyomatosis, tuberoussclerosis complex, a hamartia, or a hamartoma)), kind of concurrenttreatment, complications from the disease being treated or otherhealth-related problems. Other therapeutic regimens or agents can beused in conjunction with the methods and compounds of Applicants'invention. Adjustment and manipulation of established dosages (e.g.,frequency and duration) are well within the ability of those skilled inthe art.

For any compound described herein, the therapeutically effective amountcan be initially determined from cell culture assays. Targetconcentrations will be those concentrations of active compound(s) thatare capable of achieving the methods described herein, as measured usingthe methods described herein or known in the art.

As is well known in the art, therapeutically effective amounts for usein humans can also be determined from animal models. For example, a dosefor humans can be formulated to achieve a concentration that has beenfound to be effective in animals. The dosage in humans can be adjustedby monitoring compounds effectiveness and adjusting the dosage upwardsor downwards, as described above. Adjusting the dose to achieve maximalefficacy in humans based on the methods described above and othermethods is well within the capabilities of the ordinarily skilledartisan.

Dosages may be varied depending upon the requirements of the patient andthe compound being employed. The dose administered to a patient, in thecontext of the present invention should be sufficient to effect abeneficial therapeutic response in the patient over time. The size ofthe dose also will be determined by the existence, nature, and extent ofany adverse side-effects. Determination of the proper dosage for aparticular situation is within the skill of the practitioner. Generally,treatment is initiated with smaller dosages which are less than theoptimum dose of the compound. Thereafter, the dosage is increased bysmall increments until the optimum effect under circumstances isreached.

Dosage amounts and intervals can be adjusted individually to providelevels of the administered compound effective for the particularclinical indication being treated. This will provide a therapeuticregimen that is commensurate with the severity of the individual'sdisease state.

Utilizing the teachings provided herein, an effective prophylactic ortherapeutic treatment regimen can be planned that does not causesubstantial toxicity and yet is effective to treat the clinical symptomsdemonstrated by the particular patient. This planning should involve thecareful choice of active compound by considering factors such ascompound potency, relative bioavailability, patient body weight,presence and severity of adverse side effects, preferred mode ofadministration and the toxicity profile of the selected agent.

The compounds described herein can be used in combination with oneanother, with other active agents known to be useful in treating cancer(e.g. leukemia, lung cancer, epidermoid carcinoma (i.e. squamous-cellcarcinoma), fibrosarcoma, liver cancer (e.g. hepatocellular carcinoma),prostate cancer, kidney cancer (e.g. renal cell carcinoma or urothelialcell carcinoma), lymphoma, breast cancer, urinary bladder cancer,prostate cancer, or therapy resistant cancer) or a non-malignanthyperproliferative disease (e.g. hamartomatous lesion, angiomyolipoma,lymphangioleiomyomatosis, tuberous sclerosis complex, a hamartia, or ahamartoma), or with adjunctive agents that may not be effective alone,but may contribute to the efficacy of the active agent.

In some embodiments, co-administration includes administering one activeagent within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours of a secondactive agent. Co-administration includes administering two active agentssimultaneously, approximately simultaneously (e.g., within about 1, 5,10, 15, 20, or 30 minutes of each other), or sequentially in any order.In some embodiments, co-administration can be accomplished byco-formulation, i.e., preparing a single pharmaceutical compositionincluding both active agents. In other embodiments, the active agentscan be formulated separately. In another embodiment, the active and/oradjunctive agents may be linked or conjugated to one another. In someembodiments, the compounds described herein may be combined withtreatments for cancer (e.g. leukemia, lung cancer, epidermoid carcinoma(i.e. squamous-cell carcinoma), fibrosarcoma, liver cancer (e.g.hepatocellular carcinoma), prostate cancer, kidney cancer (e.g. renalcell carcinoma or urothelial cell carcinoma), lymphoma, breast cancer,urinary bladder cancer, prostate cancer, or therapy resistant cancer) ornon-malignant hyperproliferative diseases (e.g. hamartomatous lesion,angiomyolipoma, lymphangioleiomyomatosis, tuberous sclerosis complex, ahamartia, or a hamartoma) such as surgery.

The term “NFκB” or “NF-κB” refers to a protein complex controlling DNAtranscription commonly referred to in the art as “nuclear factorkappa-light-chain-enhancer of activated B cells” including individualproteins thereof and homologs thereof. In embodiments, “NFκB” refers tothe human protein. Included in the term “NFκB” are the wildtype andmutant forms of the protein. In embodiments, the term “NFκB” refers tothe monomeric protein. In embodiments, the term “NFκB” refers to a dimerof NFκB (e.g. homodimer or heterodimer). In some embodiments, the term“NFκB” refers to a functional complex including NFκB proteins (e.g.homodimer or heterodimer). In embodiments, the term “NFκB” refers to thehuman protein also known as NFκB1, corresponding to Entrez 4790, OMIM164011, RefSeq NM_003998, and/or UniProt P19838. In embodiments, theterm “NFκB” refers to an NFκB dimer (e.g. homodimer or heterodimer)including the human protein corresponding to Entrez 4790, OMIM 164011,RefSeq NM_003998, and/or UniProt P19838. In embodiments, the term “NFκB”refers to the human protein also known as RelA or p65, corresponding toEntrez 5970, OMIM 164014, RefSeq NM_021975, and/or UniProt Q04206. Inembodiments, the term “NFκB” refers to an NFκB dimer (e.g. homodimer orheterodimer) including the human protein corresponding to Entrez 5970,OMIM 164014, RefSeq NM_021975, and/or UniProt Q04206. In embodiments,the term “NFκB” refers to the human protein also known as NFκB2,corresponding to Entrez 4791, OMIM 164012, RefSeq NM_002502, and/orUniProt Q00653. In embodiments, the term “NFκB” refers to an NFκB dimer(e.g. homodimer or heterodimer) including the human proteincorresponding to Entrez 4791, OMIM 164012, RefSeq NM_002502, and/orUniProt Q00653. In embodiments, the term “NFκB” refers to the humanprotein also known as RelB, corresponding to Entrez 5971, OMIM 604758,RefSeq NM_006509, and/or UniProt Q01201. In embodiments, the term “NFκB”refers to an NFκB dimer (e.g. homodimer or heterodimer) including thehuman protein corresponding to Entrez 5971, OMIM 604758, RefSeqNM_006509, and/or UniProt Q01201. In embodiments, the term “NFκB” refersto the human protein also known as c-Rel, corresponding to Entrez 5966,OMIM 164910, RefSeq NM_002908, and/or UniProt Q04864. In embodiments,the term “NFκB” refers to an NFκB dimer (e.g. homodimer or heterodimer)including the human protein corresponding to Entrez 5966, OMIM 164910,RefSeq NM_002908, and/or UniProt Q04864. In embodiments, “NFκB” refersto the complex of human NFκB1 and RelA. In embodiments, “NFκB” refers tothe complex of human NFκB1 and NFκB2. In embodiments, “NFκB” refers tothe complex of human NFκB1 and RelB. In embodiments, “NFκB” refers tothe complex of human NFκB1 and c-Rel. In embodiments, “NFκB” refers tothe complex of human NFκB2 and RelA. In embodiments, “NFκB” refers tothe complex of human NFκB2 and RelB. In embodiments, “NFκB” refers tothe complex of human NFκB2 and c-Rel. In embodiments, “NFκB” refers tothe complex of human RelA and RelB. In embodiments, “NFκB” refers to thecomplex of human RelA and c-Rel. In embodiments, “NFκB” refers to thecomplex of human RelB and c-Rel. In embodiments, “NFκB” refers to thehomodimer of human NFκB1. In embodiments, “NFκB” refers to the homodimerof human NFκB2. In embodiments, “NFκB” refers to the homodimer of humanRelA. In embodiments, “NFκB” refers to the homodimer of human RelB. Inembodiments, “NFκB” refers to the homodimer of human c-Rel. Inembodiments, the reference numbers above for “NFκB” refer to theprotein, and associated nucleic acids, known as of the date of filing ofthis application. The term “TSC1” or “hamartin” refers to the protein“tuberous sclerosis protein 1”. In embodiments, “TSC1” refers to thehuman protein. Included in the term “TSC2” are the widltype and mutantforms of the protein. In embodiments, “TSC1” refers to the proteinassociated with Entrez Gene 7248, OMIM 605284, UniProt Q92574, and/orRefSeq (protein) NP_000359. In embodiments, the reference numbersimmediately above refer to the protein, and associated nucleic acids,known as of the date of filing of this application. The term “TSC2” or“tuberin” refers to the protein “tuberous sclerosis protein 2”. Inembodiments, “TSC2” refers to the human protein. Included in the term“TSC2” are the widltype and mutant forms of the protein. In embodiments,“TSC2” refers to the protein associated with Entrez Gene 7249, OMIM191092, UniProt P49815, and/or RefSeq (protein) NP_000539. Inembodiments, the reference numbers immediately above refer to theprotein, and associated nucleic acids, known as of the date of filing ofthis application. The term “mTOR” refers to the protein “mammaliantarget of rapamycin”. In embodiments, “mTOR” refers to the humanprotein. Included in the term “mTOR” are the wildtype and mutant formsof the protein. In embodiments, “mTOR” refers to the protein associatedwith Entrez Gene 2475, OMIM 601231, UniProt P42345, and/or RefSeq(protein) NP_004949. In embodiments, the reference numbers immediatelyabove refer to the protein, and associated nucleic acids, known as ofthe date of filing of this application.

“Anti-cancer agent” is used in accordance with its plain ordinarymeaning and refers to a composition (e.g. compound, drug, antagonist,inhibitor, modulator) having antineoplastic properties or the ability toinhibit the growth or proliferation of cells. In some embodiments, ananti-cancer agent is a chemotherapeutic. In some embodiments, ananti-cancer agent is an agent identified herein having utility inmethods of treating cancer. In some embodiments, an anti-cancer agent isan agent approved by the FDA or similar regulatory agency of a countryother than the USA, for treating cancer. Examples of anti-cancer agentsinclude, but are not limited to, MEK (e.g. MEK1, MEK2, or MEK1 and MEK2)inhibitors (e.g. XL518, CI-1040, PD035901, selumetinib/AZD6244,GSK1120212/trametinib, GDC-0973, ARRY-162, ARRY-300, AZD8330, PD0325901,U0126, PD98059, TAK-733, PD318088, AS703026, BAY 869766), alkylatingagents (e.g., cyclophosphamide, ifosfamide, chlorambucil, busulfan,melphalan, mechlorethamine, uramustine, thiotepa, nitrosoureas, nitrogenmustards (e.g., mechloroethamine, cyclophosphamide, chlorambucil,meiphalan), ethylenimine and methylmelamines (e.g., hexamethlymelamine,thiotepa), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g.,carmustine, lomusitne, semustine, streptozocin), triazenes(decarbazine)), anti-metabolites (e.g., 5-azathioprine, leucovorin,capecitabine, fludarabine, gemcitabine, pemetrexed, raltitrexed, folicacid analog (e.g., methotrexate), or pyrimidine analogs (e.g.,fluorouracil, floxouridine, Cytarabine), purine analogs (e.g.,mercaptopurine, thioguanine, pentostatin), etc.), plant alkaloids (e.g.,vincristine, vinblastine, vinorelbine, vindesine, podophyllotoxin,paclitaxel, docetaxel, etc.), topoisomerase inhibitors (e.g.,irinotecan, topotecan, amsacrine, etoposide (VP16), etoposide phosphate,teniposide, etc.), antitumor antibiotics (e.g., doxorubicin, adriamycin,daunorubicin, epirubicin, actinomycin, bleomycin, mitomycin,mitoxantrone, plicamycin, etc.), platinum-based compounds or platinumcontaining agents (e.g. cisplatin, oxaloplatin, carboplatin),anthracenedione (e.g., mitoxantrone), substituted urea (e.g.,hydroxyurea), methyl hydrazine derivative (e.g., procarbazine),adrenocortical suppressant (e.g., mitotane, aminoglutethimide),epipodophyllotoxins (e.g., etoposide), antibiotics (e.g., daunorubicin,doxorubicin, bleomycin), enzymes (e.g., L-asparaginase), inhibitors ofmitogen-activated protein kinase signaling (e.g. U0126, PD98059,PD184352, PD0325901, ARRY-142886, SB239063, SP600125, BAY 43-9006,wortmannin, or LY294002, Syk inhibitors, mTOR inhibitors, antibodies(e.g., rituxan), gossyphol, genasense, polyphenol E, Chlorofusin, alltrans-retinoic acid (ATRA), bryostatin, tumor necrosis factor-relatedapoptosis-inducing ligand (TRAIL), 5-aza-2′-deoxycytidine, all transretinoic acid, doxorubicin, vincristine, etoposide, gemcitabine,imatinib (Gleevec®), geldanamycin,17-N-Allylamino-17-Demethoxygeldanamycin (17-AAG), flavopiridol,LY294002, bortezomib, trastuzumab, BAY 11-7082, PKC412, PD184352,20-epi-1, 25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone;aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TKantagonists; altretamine; ambamustine; amidox; amifostine;aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole;andrographolide; angiogenesis inhibitors; antagonist D; antagonist G;antarelix; anti-dorsalizing morphogenetic protein-1; antiandrogen,prostatic carcinoma; antiestrogen; antineoplaston; antisenseoligonucleotides; aphidicolin glycinate; apoptosis gene modulators;apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA; argininedeaminase; asulacrine; atamestane; atrimustine; axinastatin 1;axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatinIII derivatives; balanol; batimastat; BCR/ABL antagonists;benzochlorins; benzoylstaurosporine; beta lactam derivatives;beta-alethine; betaclamycin B; betulinic acid; bFGF inhibitor;bicalutamide; bisantrene; bisaziridinylspermine; bisnafide; bistrateneA; bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine;calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2;capecitabine; carboxamide-amino-triazole; carboxyamidotriazole; CaRestM3; CARN 700; cartilage derived inhibitor; carzelesin; casein kinaseinhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorins;chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; cladribine;clomifene analogues; clotrimazole; collismycin A; collismycin B;combretastatin A4; combretastatin analogue; conagenin; crambescidin 816;crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A;cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate;cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin B;deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil;diaziquone; didemnin B; didox; diethylnorspermine;dihydro-5-azacytidine; 9-dioxamycin; diphenyl spiromustine; docosanol;dolasetron; doxifluridine; droloxifene; dronabinol; duocarmycin SA;ebselen; ecomustine; edelfosine; edrecolomab; eflornithine; elemene;emitefur; epirubicin; epristeride; estramustine analogue; estrogenagonists; estrogen antagonists; etanidazole; etoposide phosphate;exemestane; fadrozole; fazarabine; fenretinide; filgrastim; finasteride;flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicinhydrochloride; forfenimex; formestane; fostriecin; fotemustine;gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix;gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam;heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid;idarubicin; idoxifene; idramantone; ilmofosine; ilomastat;imidazoacridones; imiquimod; immunostimulant peptides; insulin-likegrowth factor-1 receptor inhibitor; interferon agonists; interferons;interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact;irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine;lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides;maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysininhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone;meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone;miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone;mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growthfactor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonalantibody, human chorionic gonadotrophin; monophosphoryl lipidA+myobacterium cell wall sk; mopidamol; multiple drug resistance geneinhibitor; multiple tumor suppressor 1-based therapy; mustard anticanceragent; mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; neutral endopeptidase; nilutamide;nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn;O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; palauamine; palmitoylrhizoxin;pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine;pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin;pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylerie conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide;rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol;saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics;semustine; senescence derived inhibitor 1; sense oligonucleotides;signal transduction inhibitors; signal transduction modulators; singlechain antigen-binding protein; sizofuran; sobuzoxane; sodiumborocaptate; sodium phenylacetate; solverol; somatomedin bindingprotein; sonermin; sparfosic acid; spicamycin D; spiromustine;splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-celldivision inhibitors; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; synthetic glycosaminoglycans; tallimustine;tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium;tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; totipotent stem cell factor;translation inhibitors; tretinoin; triacetyluridine; triciribine;trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinaseinhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenitalsinus-derived growth inhibitory factor; urokinase receptor antagonists;vapreotide; variolin B; vector system, erythrocyte gene therapy;velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine;vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; zinostatinstimalamer, Adriamycin, Dactinomycin, Bleomycin, Vinblastine, Cisplatin,acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin;aldesleukin; altretamine; ambomycin; ametantrone acetate;aminoglutethimide; amsacrine; anastrozole; anthramycin; asparaginase;asperlin; azacitidine; azetepa; azotomycin; batimastat; benzodepa;bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin;bleomycin sulfate; brequinar sodium; bropirimine; busulfan;cactinomycin; calusterone; caracemide; carbetimer; carboplatin;carmustine; carubicin hydrochloride; carzelesin; cedefingol;chlorambucil; cirolemycin; cladribine; crisnatol mesylate;cyclophosphamide; cytarabine; dacarbazine; daunorubicin hydrochloride;decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate;diaziquone; doxorubicin; doxorubicin hydrochloride; droloxifene;droloxifene citrate; dromostanolone propionate; duazomycin; edatrexate;eflornithine hydrochloride; elsamitrucin; enloplatin; enpromate;epipropidine; epirubicin hydrochloride; erbulozole; esorubicinhydrochloride; estramustine; estramustine phosphate sodium; etanidazole;etoposide; etoposide phosphate; etoprine; fadrozole hydrochloride;fazarabine; fenretinide; floxuridine; fludarabine phosphate;fluorouracil; fluorocitabine; fosquidone; fostriecin sodium;gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicinhydrochloride; ifosfamide; iimofosine; interleukin I1 (includingrecombinant interleukin II, or rlL.sub.2), interferon alfa-2a;interferon alfa-2b; interferon alfa-n1; interferon alfa-n3; interferonbeta-1a; interferon gamma-1b; iproplatin; irinotecan hydrochloride;lanreotide acetate; letrozole; leuprolide acetate; liarozolehydrochloride; lometrexol sodium; lomustine; losoxantrone hydrochloride;masoprocol; maytansine; mechlorethamine hydrochloride; megestrolacetate; melengestrol acetate; melphalan; menogaril; mercaptopurine;methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide;mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper;mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazoie;nogalamycin; ormaplatin; oxisuran; pegaspargase; peliomycin;pentamustine; peplomycin sulfate; perfosfamide; pipobroman; piposulfan;piroxantrone hydrochloride; plicamycin; plomestane; porfimer sodium;porfiromycin; prednimustine; procarbazine hydrochloride; puromycin;puromycin hydrochloride; pyrazofurin; riboprine; rogletimide; safingol;safingol hydrochloride; semustine; simtrazene; sparfosate sodium;sparsomycin; spirogermanium hydrochloride; spiromustine; spiroplatin;streptonigrin; streptozocin; sulofenur; talisomycin; tecogalan sodium;tegafur; teloxantrone hydrochloride; temoporfin; teniposide; teroxirone;testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;tirapazamine; toremifene citrate; trestolone acetate; triciribinephosphate; trimetrexate; trimetrexate glucuronate; triptorelin;tubulozole hydrochloride; uracil mustard; uredepa; vapreotide;verteporfin; vinblastine sulfate; vincristine sulfate; vindesine;vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicinhydrochloride, agents that arrest cells in the G2-M phases and/ormodulate the formation or stability of microtubules, (e.g. Taxol™ (i.e.paclitaxel), Taxotere™, compounds comprising the taxane skeleton,Erbulozole (i.e. R-55104), Dolastatin 10 (i.e. DLS-10 and NSC-376128),Mivobulin isethionate (i.e. as CI-980), Vincristine, NSC-639829,Discodermolide (i.e. as NVP-XX-A-296), ABT-751 (Abbott, i.e. E-7010),Altorhyrtins (e.g. Altorhyrtin A and Altorhyrtin C), Spongistatins (e.g.Spongistatin 1, Spongistatin 2, Spongistatin 3, Spongistatin 4,Spongistatin 5, Spongistatin 6, Spongistatin 7, Spongistatin 8, andSpongistatin 9), Cemadotin hydrochloride (i.e. LU-103793 andNSC-D-669356), Epothilones (e.g. Epothilone A, Epothilone B, EpothiloneC (i.e. desoxyepothilone A or dEpoA), Epothilone D (i.e. KOS-862, dEpoB,and desoxyepothilone B), Epothilone E, Epothilone F, Epothilone BN-oxide, Epothilone A N-oxide, 16-aza-epothilone B, 21-aminoepothilone B(i.e. BMS-310705), 21-hydroxyepothilone D (i.e. Desoxyepothilone F anddEpoF), 26-fluoroepothilone, Auristatin PE (i.e. NSC-654663), Soblidotin(i.e. TZT-1027), LS-4559-P (Pharmacia, i.e. LS-4577), LS-4578(Pharmacia, i.e. LS-477-P), LS-4477 (Pharmacia), LS-4559 (Pharmacia),RPR-112378 (Aventis), Vincristine sulfate, DZ-3358 (Daiichi), FR-182877(Fujisawa, i.e. WS-9885B), GS-164 (Takeda), GS-198 (Takeda), KAR-2(Hungarian Academy of Sciences), BSF-223651 (BASF, i.e. ILX-651 andLU-223651), SAH-49960 (Lilly/Novartis), SDZ-268970 (Lilly/Novartis),AM-97 (Armad/Kyowa Hakko), AM-132 (Armad), AM-138 (Armad/Kyowa Hakko),IDN-5005 (Indena), Cryptophycin 52 (i.e. LY-355703), AC-7739 (Ajinomoto,i.e. AVE-8063A and CS-39.HCl), AC-7700 (Ajinomoto, i.e. AVE-8062,AVE-8062A, CS-39-L-Ser.HCl, and RPR-258062A), Vitilevuamide, TubulysinA, Canadensol, Centaureidin (i.e. NSC-106969), T-138067 (Tularik, i.e.T-67, TL-138067 and TI-138067), COBRA-1 (Parker Hughes Institute, i.e.DDE-261 and WHI-261), H10 (Kansas State University), H16 (Kansas StateUniversity), Oncocidin A1 (i.e. BTO-956 and DIME), DDE-313 (ParkerHughes Institute), Fijianolide B, Laulimalide, SPA-2 (Parker HughesInstitute), SPA-1 (Parker Hughes Institute, i.e. SPIKET-P), 3-IAABU(Cytoskeleton/Mt. Sinai School of Medicine, i.e. MF-569), Narcosine(also known as NSC-5366), Nascapine, D-24851 (Asta Medica), A-105972(Abbott), Hemiasterlin, 3-BAABU (Cytoskeleton/Mt. Sinai School ofMedicine, i.e. MF-191), TMPN (Arizona State University), Vanadoceneacetylacetonate, T-138026 (Tularik), Monsatrol, lnanocine (i.e.NSC-698666), 3-IAABE (Cytoskeleton/Mt. Sinai School of Medicine),A-204197 (Abbott), T-607 (Tuiarik, i.e. T-900607), RPR-115781 (Aventis),Eleutherobins (such as Desmethyleleutherobin, Desaetyleleutherobin,lsoeleutherobin A, and Z-Eleutherobin), Caribaeoside, Caribaeolin,Halichondrin B, D-64131 (Asta Medica), D-68144 (Asta Medica),Diazonamide A, A-293620 (Abbott), NPI-2350 (Nereus), Taccalonolide A,TUB-245 (Aventis), A-259754 (Abbott), Diozostatin, (−)-Phenylahistin(i.e. NSCL-96F037), D-68838 (Asta Medica), D-68836 (Asta Medica),Myoseverin B, D-43411 (Zentaris, i.e. D-81862), A-289099 (Abbott),A-318315 (Abbott), HTI-286 (i.e. SPA-110, trifluoroacetate salt)(Wyeth), D-82317 (Zentaris), D-82318 (Zentaris), SC-12983 (NCI),Resverastatin phosphate sodium, BPR-OY-007 (National Health ResearchInstitutes), and SSR-250411 (Sanofi)), steroids (e.g., dexamethasone),finasteride, aromatase inhibitors, gonadotropin-releasing hormoneagonists (GnRH) such as goserelin or leuprolide, adrenocorticosteroids(e.g., prednisone), progestins (e.g., hydroxyprogesterone caproate,megestrol acetate, medroxyprogesterone acetate), estrogens (e.g.,diethlystilbestrol, ethinyl estradiol), antiestrogen (e.g., tamoxifen),androgens (e.g., testosterone propionate, fluoxymesterone), antiandrogen(e.g., flutamide), immunostimulants (e.g., Bacillus Calmette-Guérin(BCG), levamisole, interleukin-2, alpha-interferon, etc.), monoclonalantibodies (e.g., anti-CD20, anti-HER2, anti-CD52, anti-HLA-DR, andanti-VEGF monoclonal antibodies), immunotoxins (e.g., anti-CD33monoclonal antibody-calicheamicin conjugate, anti-CD22 monoclonalantibody-pseudomonas exotoxin conjugate, etc.), radioimmunotherapy(e.g., anti-CD20 monoclonal antibody conjugated to ¹¹¹In, ⁹⁰Y, or ¹³¹I,etc.), triptolide, homoharringtonine, dactinomycin, doxorubicin,epirubicin, topotecan, itraconazole, vindesine, cerivastatin,vincristine, deoxyadenosine, sertraline, pitavastatin, irinotecan,clofazimine, 5-nonyloxytryptamine, vemurafenib, dabrafenib, erlotinib,gefitinib, EGFR inhibitors, epidermal growth factor receptor(EGFR)-targeted therapy or therapeutic (e.g. gefitinib (Iressa™)erlotinib (Tarceva™) cetuximab (Erbitux™), lapatinib (Tykerb™),panitumumab (Vectibix™) vandetanib (Caprelsa™), afatinib/BIBW2992,CI-1033/canertinib, neratinib/HKI-272, CP-724714, TAK-285, AST-1306,ARRY334543, ARRY-380, AG-1478, dacomitinib/PF299804, OSI-420/desmethylerlotinib, AZD8931, AEE788, pelitinib/EKB-569, CUDC-101, WZ8040, WZ4002,WZ3146, AG-490, XL647, PD153035, BMS-599626), sorafenib, imatinib,sunitinib, dasatinib, hormonal therapies, or the like.

The term “platinum-based compound” or “platinum containing agent” asused herein refers to a compound comprising a heavy metal complexcontaining a central atom of platinum surrounded by organic and/orinorganic functionalities. Non-limiting examples of platinum-basedcompounds include oxaliplatin, cisplatin, carboplatin, pharmaceuticallyacceptable salts thereof, stereoisomers thereof, derivatives thereof,analogs thereof, and combinations thereof. Included withinplatinum-based compounds are platinum-based drugs.

“Hormonal therapy” or “hormone therapy” is used in accordance with itsplain ordinary meaning and refers to the treatment of a disease (e.g.cancer) by administration of an agent (e.g. compound) that modulates theproduction or activity of a hormone, the activity of a protein that ismodulated by a hormone, or a signaling pathway that is modulated by ahormone or the activity of a hormone. An agent used in hormonal therapymay be refered to as a “hormonal therapy agent” or “hormonal therapeuticagent”. Hormonal therapeutic agents include, but are not limited to,estrogens, androgens, antiestrogens, antiandrogens, endocrine therapies,steroids (e.g., dexamethasone), finasteride, fulvestrant, aromataseinhibitors (e.g. exemestane, anastrozole, aminoglutethimide,testolactone, letrozole, vorozole, formestane, or fadrozole), tamoxifen,and gonadotropin-releasing hormone agonists (GnRH) such as goserelin.

“Chemotherapeutic” or “chemotherapeutic agent” is used in accordancewith its plain ordinary meaning and refers to a chemical composition orcompound having antineoplastic properties or the ability to inhibit thegrowth or proliferation of cells.

“Therapy resistant” cancers, tumor cells, and tumors refer to cancersthat have become resistant to one or more cancer therapies including,but not limited to, an anti-cancer agent, a chemotherapy (e.g. achemotherapeutic), a hormonal therapy (e.g. a hormonal therapeuticagent), a radiotherapy, an immunotherapy, and/or combinations thereof.

Additionally, the compounds described herein can be co-administered withconventional immunotherapeutic agents including, but not limited to,immunostimulants (e.g., Bacillus Calmette-Guérin (BCG), levamisole,interleukin-2, alpha-interferon, etc.), monoclonal antibodies (e.g.,anti-CD20, anti-HER2, anti-CD52, anti-HLA-DR, and anti-VEGF monoclonalantibodies), immunotoxins (e.g., anti-CD33 monoclonalantibody-calicheamicin conjugate, anti-CD22 monoclonalantibody-pseudomonas exotoxin conjugate, etc.), and radioimmunotherapy(e.g., anti-CD20 monoclonal antibody conjugated to ¹¹¹In, ⁹⁰Y or ¹³¹Ietc.).

In a further embodiment, the compounds described herein can beco-administered with conventional radiotherapeutic agents including, butnot limited to, radionuclides such as ⁴⁷Sc, ⁶⁴Cu, ⁶⁷Cu, ⁸⁹Sr, ⁸⁶Y, ⁸⁷Y,⁹⁰Y, ¹⁰⁵Rh, ¹¹¹Ag, ^(117m)Sn, ¹⁴⁹Pm, ¹⁵³Sm, ¹⁶⁶Ho, ¹⁷⁷Lu, ¹⁸⁶Re, ¹⁸⁸Re,²¹¹At, and ²¹²Bi, optionally conjugated to antibodies directed againsttumor antigens.

The term “cancer stem cell” is used in accordance with its plainordinary meaning within cancer biology and cell biology and refers tocancer cells with characteristics associated with non-cancerous stemcells, including for example the ability of self-renewal and the abilityto differentiate into all cell types that make up a cancer sample (e.g.tumor or cancer cells). The term “cancer progenitor cell” is used inaccordance with its plain ordinary meaning within cancer biology andcell biology and refers to cancer cells with characteristics associatedwith non-cancerous progenitor cells. In embodiments, cancer stem cellsand/or cancer progenitor cells may be identified by the presence (orabsence) of cell markers (e.g. CD133 (PROM1), CD44 (PGP1), CD24 (HSA),EpCAM (epithelial cell adhesion molecule, ESA (epithelial specificantigen)), THY1 (CD90), ATP-binding cassette B5 (ABCBS), Hoechst33342,CD34, and/or ALDH1 (aldehyde dehydrogenase). In embodiments, lung cancerstem/progenitor cells may be identified by the presence of CD133 andALDH1 (e.g. high levels of ALDH1 compared to non-lung cancerstem/progenitor cells).

A “stem cell” is a cell characterized by the ability of self-renewalthrough mitotic cell division and the potential to differentiate into atissue or an organ. Among mammalian stem cells, embryonic and somaticstem cells can be distinguished. Embryonic stem cells reside in theblastocyst and give rise to embryonic tissues, whereas somatic stemcells reside in adult tissues for the purpose of tissue regeneration andrepair.

“Self renewal” refers to the ability of a cell to divide and generate atleast one daughter cell with the self-renewing characteristics of theparent cell. The second daughter cell may commit to a particulardifferentiation pathway. For example, a self-renewing hematopoietic stemcell can divide and form one daughter stem cell and another daughtercell committed to differentiation in the myeloid or lymphoid pathway. Acommitted progenitor cell has typically lost the self-renewal capacity,and upon cell division produces two daughter cells that display a moredifferentiated (i.e., restricted) phenotype. Non-self renewing cellsrefers to cells that undergo cell division to produce daughter cells,neither of which have the differentiation potential of the parent celltype, but instead generates differentiated daughter cells.

The term “pluripotent” or “pluripotency” refers to cells with theability to give rise to progeny that can undergo differentiation, underappropriate conditions, into cell types that collectively exhibitcharacteristics associated with cell lineages from the three germ layers(endoderm, mesoderm, and ectoderm). Pluripotent stem cells cancontribute to tissues of a prenatal, postnatal or adult organism. Astandard art-accepted test, such as the ability to form a teratoma in8-12 week old SCID mice, can be used to establish the pluripotency of acell population. However, identification of various pluripotent stemcell characteristics can also be used to identify pluripotent cells.

“Pluripotent stem cell characteristics” refer to characteristics of acell that distinguish pluripotent stem cells from other cells.Expression or non-expression of certain combinations of molecularmarkers are examples of characteristics of pluripotent stem cells. Morespecifically, human pluripotent stem cells may express at least some,and optionally all, of the markers from the following non-limiting list:SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, TRA-2-49/6E, ALP, Sox2, E-cadherin,UTF-1, Oct4, Lin28, Rex1, and Nanog. Cell morphologies associated withpluripotent stem cells are also pluripotent stem cell characteristics.

The terms “induced pluripotent stem cell,” “iPS” and the like refer to apluripotent stem cell artificially derived from a non-pluripotent cell.A “non-pluripotent cell” can be a cell of lesser potency to self-renewand differentiate than a pluripotent stem cell. Cells of lesser potencycan be, but are not limited to adult stem cells, tissue specificprogenitor cells, primary or secondary cells.

An adult stem cell is an undifferentiated cell found in an individualafter embryonic development. Adult stem cells multiply by cell divisionto replenish dying cells and regenerate damaged tissue. An adult stemcell has the ability to divide and create another cell like itself or tocreate a more differentiated cell. Even though adult stem cells areassociated with the expression of pluripotency markers such as Rex1,Nanog, Oct4 or Sox2, they do not have the ability of pluripotent stemcells to differentiate into the cell types of all three germ layers.Adult stem cells have a limited ability to self renew and generateprogeny of distinct cell types. Adult stem cells can includehematopoietic stem cell, a cord blood stem cell, a mesenchymal stemcell, an epithelial stem cell, a skin stem cell or a neural stem cell. Atissue specific progenitor refers to a cell devoid of self-renewalpotential that is committed to differentiate into a specific organ ortissue. A primary cell includes any cell of an adult or fetal organismapart from egg cells, sperm cells and stem cells. Examples of usefulprimary cells include, but are not limited to, skin cells, bone cells,blood cells, cells of internal organs and cells of connective tissue. Asecondary cell is derived from a primary cell and has been immortalizedfor long-lived in vitro cell culture.

II. Compounds

In a first aspect is provided a compound, or a pharmaceuticallyacceptable salt thereof, wherein the compound has a formula selectedfrom the group consisting of:

L⁰ is independently a bond or an unsubstituted C₁-C₁₀ alkylene. R¹ andR² are independently hydrogen, —OH, substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, or substituted or unsubstituted heteroaryl; whereR¹ and R² may optionally be joined to form a substituted orunsubstituted heterocycloalkyl or a substituted or unsubstitutedheteroaryl.

In embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the formula:

In embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the formula:

In embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the formula:

In embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the formula:

In embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the formula:

In embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the formula:

In embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the formula:

In embodiments, the compound, or a pharmaceutically acceptable saltthereof, has the formula:

In embodiments, L⁰ is a bond. In embodiments, L⁰ is an unsubstitutedC₁-C₆ alkylene. In embodiments, L⁰ is an unsubstituted C₁-C₄ alkylene.In embodiments, L⁰ is an unsubstituted methylene.

In embodiments, R¹ is L¹-R³; L¹ is independently a bond or anunsubstituted C₁-C₁₀ alkylene; and R³ is independently hydrogen,halogen, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂,—NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂,—NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H,—NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, or substituted or unsubstitutedheteroaryl. In embodiments, L¹ is a bond. In embodiments, L¹ is anunsubstituted C₁-C₄ alkylene. In embodiments, L¹ is an unsubstitutedmethylene.

In embodiments, R³ is independently hydrogen, halogen, —CH₃, —CF₃,—CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃,—CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃,—OCF₃, —OCHF₂, 4-methoxybenzyl, 4-methoxyphenyl, or pyridyl.

In embodiments, R² is L²-R⁴; L² is independently a bond or anunsubstituted C₁-C₁₀ alkylene; and R⁴ is independently hydrogen,halogen, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂,—NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂,—NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H,—NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, or substituted or unsubstitutedheteroaryl. In embodiments, L² is a bond. In embodiments, L² is anunsubstituted C₁-C₄ alkylene. In embodiments, L² is an unsubstitutedmethylene.

In embodiments, R⁴ is independently hydrogen, halogen, —CH₃, —CF₃,—CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃,—CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃,—OCF₃, —OCHF₂, 4-methoxybenzyl, 4-methoxyphenyl, or pyridyl.

In embodiments, R¹ and R² are joined to form an unsubstitutedheterocycloalkyl. In embodiments, R¹ and R² are joined to form anunsubstituted piperidinyl.

In embodiments of the pharmaceutically acceptable salt, the compound hasa formula selected from the group consisting of formula I, II, IV, Ia,IIa, and IVa, wherein the compound includes a protonated nitrogencation. In embodiments, the pharmaceutically acceptable salt of acompound described herein includes fumarate. In embodiments, thepharmaceutically acceptable salt of a compound described herein includesfumaric acid. In embodiments, the pharmaceutically acceptable salt of acompound described herein includes fumarate with a single negativecharge. In embodiments, the pharmaceutically acceptable salt of acompound described herein includes chloride. In embodiments, thepharmaceutically acceptable salt of a compound described herein includesbromide. In embodiments, the pharmaceutically acceptable salt of acompound described herein includes sulfate. In embodiments, thepharmaceutically acceptable salt of a compound described herein includesmethanesulfonate. In embodiments, the pharmaceutically acceptable saltof a compound described herein includes nitrate. In embodiments, thepharmaceutically acceptable salt of a compound described herein includesmaleate. In embodiments, the pharmaceutically acceptable salt of acompound described herein includes acetate. In embodiments, thepharmaceutically acceptable salt of a compound described herein includescitrate. In embodiments, the pharmaceutically acceptable salt of acompound described herein includes tartrate. In embodiments, thepharmaceutically acceptable salt of a compound described herein includessuccinate. In embodiments, the pharmaceutically acceptable salt of acompound described herein includes benzoate. In embodiments, thepharmaceutically acceptable salt of a compound described herein includesglutamate.

In embodiments, R¹ is independently hydrogen, R¹¹-substituted orunsubstituted alkyl, R¹¹-substituted or unsubstituted heteroalkyl,R¹¹-substituted or unsubstituted cycloalkyl, R¹¹-substituted orunsubstituted heterocycloalkyl, R¹¹-substituted or unsubstituted aryl,or R¹¹-substituted or unsubstituted heteroaryl. In embodiments, R¹ andR² may optionally be joined to form an R¹¹-substituted or unsubstitutedheterocycloalkyl or R¹¹-substituted or unsubstituted heteroaryl.

R¹¹ is independently hydrogen, halogen, oxo, —CH₃, —CF₃, —CCl₃, —CN,—S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH,—SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,R¹²-substituted or unsubstituted alkyl, R¹²-substituted or unsubstitutedheteroalkyl, R¹²-substituted or unsubstituted cycloalkyl,R¹²-substituted or unsubstituted heterocycloalkyl, R¹²-substituted orunsubstituted aryl, or R¹²-substituted or unsubstituted heteroaryl.

In embodiments, R² is independently hydrogen, halogen, —CH₃, —CF₃,—CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃,—CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃,—OCF₃, —OCHF₂, R¹³-substituted or unsubstituted alkyl, R¹³-substitutedor unsubstituted heteroalkyl, R¹³-substituted or unsubstitutedcycloalkyl, R¹³-substituted or unsubstituted heterocycloalkyl,R¹³-substituted or unsubstituted aryl, or R¹³-substituted orunsubstituted heteroaryl. In embodiments, R¹ and R² may optionally bejoined to form an R¹³-substituted or unsubstituted heterocycloalkyl orR¹³-substituted or unsubstituted heteroaryl.

R¹³ is independently hydrogen, halogen, oxo, —CH₃, —CF₃, —CCl₃, —CN,—S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH,—SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,R¹⁴-substituted or unsubstituted alkyl, R¹⁴-substituted or unsubstitutedheteroalkyl, R¹⁴-substituted or unsubstituted cycloalkyl,R¹⁴-substituted or unsubstituted heterocycloalkyl, R¹⁴-substituted orunsubstituted aryl, or R¹⁴-substituted or unsubstituted heteroaryl.

In embodiments, R³ is independently hydrogen, halogen, —CH₃, —CF₃,—CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃,—CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃,—OCF₃, —OCHF₂, R¹⁵-substituted or unsubstituted alkyl, R¹⁵-substitutedor unsubstituted heteroalkyl, R¹⁵-substituted or unsubstitutedcycloalkyl, R¹⁵-substituted or unsubstituted heterocycloalkyl,R¹⁵-substituted or unsubstituted aryl, or R¹⁵-substituted orunsubstituted heteroaryl.

R¹⁵ is independently hydrogen, halogen, oxo, —CH₃, —CF₃, —CCl₃, —CN,—S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH,—SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,R¹⁶-substituted or unsubstituted alkyl, R¹⁶-substituted or unsubstitutedheteroalkyl, R¹⁶-substituted or unsubstituted cycloalkyl,R¹⁶-substituted or unsubstituted heterocycloalkyl, R¹⁶-substituted orunsubstituted aryl, or R¹⁶-substituted or unsubstituted heteroaryl.

In embodiments, R⁴ is independently hydrogen, halogen, —CH₃, —CF₃,—CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃,—CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃,—OCF₃, —OCHF₂, R¹⁷-substituted or unsubstituted alkyl, R¹⁷-substitutedor unsubstituted heteroalkyl, R¹⁷-substituted or unsubstitutedcycloalkyl, R¹⁷-substituted or unsubstituted heterocycloalkyl,R¹⁷-substituted or unsubstituted aryl, or R¹⁷-substituted orunsubstituted heteroaryl.

R¹⁷ is independently hydrogen, halogen, oxo, —CH₃, —CF₃, —CCl₃, —CN,—S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH,—SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,R¹⁸-substituted or unsubstituted alkyl, R¹⁸-substituted or unsubstitutedheteroalkyl, R¹⁸-substituted or unsubstituted cycloalkyl,R¹⁸-substituted or unsubstituted heterocycloalkyl, R¹⁸-substituted orunsubstituted aryl, or R¹⁸-substituted or unsubstituted heteroaryl.

Each R¹², R¹⁴, R¹⁶, and R¹⁸ is independently hydrogen, halogen, oxo,—CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂,—C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂,—NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H,—NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂, unsubstituted alkyl,unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstitutedheterocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl.

In some embodiments, a compound as described herein may include multipleinstances of R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, and/or othervariables. In such embodiments, each variable may optional be differentand be appropriately labeled to distinguish each group for greaterclarity. For example, where each R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷,and/or R¹⁸ is different, they may be referred to, for example, asR^(11.1), R^(11.2), R^(11.3), R^(11.4), R^(12.1), R^(12.2), R^(12.3),R^(12.4), R^(13.1), R^(13.2), R^(13.3), R^(13.4), R^(14.1), R^(14.2),R^(14.3), R^(14.4), R^(15.1), R^(15.2), R^(15.3), R^(15.4), R^(16.1),R^(16.2), R^(16.3), R^(16.4), R^(17.1), R^(17.2), R^(17.3), R^(17.4),R^(18.1), R^(18.2), R^(18.3), and/or R^(18.4), respectively, wherein thedefinition of R¹¹ is assumed by R^(11.1), R^(11.2), R^(11.3), and/orR^(11.4), the definition of R¹² is assumed by R^(12.1), R^(12.2),R^(12.3), and/or R^(12.4), the definition of R¹³ is assumed by R^(13.1),R^(13.12), R^(13.3), and/or R^(13.4), the definition of R¹⁴ is assumedby R^(14.1), R^(14.2), R^(14.3), and/or R^(14.4), the definition of R¹⁵is assumed by R^(15.1), R^(15.2), R^(15.3), and/or R^(15.4), thedefinition of R¹⁶ is assumed by R^(16.1), R^(16.2), R^(16.3), and/orR^(16.4), the definition of R¹⁷ is assumed by R^(17.1), R^(17.2),R^(17.3), and/or R^(17.4), and the definition of R¹⁸ is assumed byR^(18.1), R^(18.2), R^(18.3), and/or R^(18.4). The variables used withina definition of R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, and/or othervariables that appear at multiple instances and are different maysimilarly be appropriately labeled to distinguish each group for greaterclarity.

In embodiments, the compound has formula (I) and L⁰ is unsubstitutedC₁-C₁₀ alkylene. In embodiments, the compound has formula (I) and L⁰ isunsubstituted C₁-C₈ alkylene. In embodiments, the compound has formula(I) and L⁰ is unsubstituted C₁-C₆ alkylene. In embodiments, the compoundhas formula (I) and L⁰ is unsubstituted C₁-C₄ alkylene. In embodiments,the compound has formula (I) and L⁰ is unsubstituted C₁-C₃ alkylene. Inembodiments, the compound has formula (I) and L⁰ is unsubstituted C₁-C₂alkylene. In embodiments, the compound has formula (I) and L⁰ isunsubstituted branched C₁-C₄ alkylene. In embodiments, the compound hasformula (I) and L⁰ is unsubstituted branched C₁-C₃ alkylene. Inembodiments, the compound has formula (I) and L⁰ is unsubstitutedmethylene. In embodiments, the compound has formula (I) and L⁰ isunsubstituted ethylene. In embodiments, the compound has formula (I) andL⁰ is a bond.

In embodiments, the compound has formula (I) and R¹ is independentlyhydrogen. In embodiments, the compound has formula (I) and R¹ isindependently halogen. In embodiments, the compound has formula (I) andR¹ is independently —CH₃. In embodiments, the compound has formula (I)and R¹ is independently —CF₃. In embodiments, the compound has formula(I) and R¹ is independently —OH. In embodiments, the compound hasformula (I) and R¹ is independently substituted 4-methoxybenzyl. Inembodiments, the compound has formula (I) and R¹ is independentlysubstituted 4-methoxyphenyl. In embodiments, the compound has formula(I) and R¹ is independently substituted pyridyl. In embodiments, thecompound has formula (I) and R¹ is independently unsubstituted4-methoxybenzyl. In embodiments, the compound has formula (I) and R¹ isindependently unsubstituted 4-methoxyphenyl. In embodiments, thecompound has formula (I) and R¹ is independently unsubstituted pyridyl.In embodiments, the compound has formula (I) and R¹ is substituted orunsubstituted alkyl. In embodiments, the compound has formula (I) and R¹is substituted or unsubstituted heteroalkyl. In embodiments, thecompound has formula (I) and R¹ is substituted or unsubstitutedcycloalkyl. In embodiments, the compound has formula (I) and R¹ issubstituted or unsubstituted heterocycloalkyl. In embodiments, thecompound has formula (I) and R¹ is substituted or unsubstituted aryl. Inembodiments, the compound has formula (I) and R¹ is substituted orunsubstituted heteroaryl. In embodiments, the compound has formula (I)and R¹ is unsubstituted alkyl. In embodiments, the compound has formula(I) and R¹ is unsubstituted heteroalkyl. In embodiments, the compoundhas formula (I) and R¹ is unsubstituted cycloalkyl. In embodiments, thecompound has formula (I) and R¹ is unsubstituted heterocycloalkyl. Inembodiments, the compound has formula (I) and R¹ is unsubstituted aryl.In embodiments, the compound has formula (I) and R¹ is unsubstitutedheteroaryl. In embodiments, the compound has formula (I) and R¹ issubstituted or unsubstituted C₁-C₈ alkyl. In embodiments, the compoundhas formula (I) and R¹ is substituted or unsubstituted C₁-C₆ alkyl. Inembodiments, the compound has formula (I) and R¹ is substituted orunsubstituted C₁-C₄ alkyl. In embodiments, the compound has formula (I)and R¹ is substituted or unsubstituted C₁-C₂ alkyl. In embodiments, thecompound has formula (I) and R¹ is substituted or unsubstituted methyl.In embodiments, the compound has formula (I) and R¹ is unsubstitutedmethyl. In embodiments, the compound has formula (I) and R¹ issubstituted or unsubstituted 2 to 8 membered heteroalkyl. Inembodiments, the compound has formula (I) and R¹ is substituted orunsubstituted 2 to 6 membered heteroalkyl. In embodiments, the compoundhas formula (I) and R¹ is substituted or unsubstituted 2 to 4 memberedheteroalkyl. In embodiments, the compound has formula (I) and R¹ issubstituted or unsubstituted 2 to 3 membered heteroalkyl. Inembodiments, the compound has formula (I) and R¹ is substituted orunsubstituted C₃-C₇ cycloalkyl. In embodiments, the compound has formula(I) and R¹ is substituted or unsubstituted 3 to 7 memberedheterocycloalkyl. In embodiments, the compound has formula (I) and R¹ issubstituted or unsubstituted 5 to 7 membered heterocycloalkyl. Inembodiments, the compound has formula (I) and R¹ is substituted orunsubstituted 5 to 6 membered heterocycloalkyl. In embodiments, thecompound has formula (I) and R¹ is substituted or unsubstituted C₆-C₁₀aryl. In embodiments, the compound has formula (I) and R¹ is substitutedor unsubstituted phenyl. In embodiments, the compound has formula (I)and R¹ is substituted or unsubstituted 5 to 9 membered heteroaryl. Inembodiments, the compound has formula (I) and R¹ is substituted orunsubstituted 5 to 6 membered heteroaryl. In embodiments, the compoundhas formula (I) and R¹ is substituted or unsubstituted 5 memberedheteroaryl. In embodiments, the compound has formula (I) and R¹ issubstituted or unsubstituted 6 membered heteroaryl. In embodiments, thecompound has formula (I) and R¹ is substituted or unsubstituted fusedring heteroaryl. In embodiments, the compound has formula (I) and R¹ issubstituted or unsubstituted 9 membered heteroaryl. In embodiments, thecompound has formula (I) and R¹ is substituted or unsubstituted 10membered heteroaryl. In embodiments, the compound has formula (I) and R¹is —OCH₃. In embodiments, the compound has formula (I) and R¹ is—CH₂CH₂OH. In embodiments, the compound has formula (I) and R¹ is—CH₂CH₃.

In embodiments, the compound has formula (I) and R¹ is L¹-R³. Inembodiments, the compound has formula (I) and L¹ is unsubstituted C₁-C₁₀alkylene. In embodiments, the compound has formula (I) and L¹ isunsubstituted C₁-C₈ alkylene. In embodiments, the compound has formula(I) and L¹ is unsubstituted C₁-C₆ alkylene. In embodiments, the compoundhas formula (I) and L¹ is unsubstituted C₁-C₄ alkylene. In embodiments,the compound has formula (I) and L¹ is unsubstituted C₁-C₃ alkylene. Inembodiments, the compound has formula (I) and L¹ is unsubstituted C₁-C₂alkylene. In embodiments, the compound has formula (I) and L¹ isunsubstituted branched C₁-C₄ alkylene. In embodiments, the compound hasformula (I) and L¹ is unsubstituted branched C₁-C₃ alkylene. Inembodiments, the compound has formula (I) and L¹ is unsubstitutedmethylene. In embodiments, the compound has formula (I) and L¹ isunsubstituted ethylene. In embodiments, the compound has formula (I) andL¹ is a bond. In embodiments, the compound has formula (I) and R³ isindependently hydrogen. In embodiments, the compound has formula (I) andR³ is independently halogen. In embodiments, the compound has formula(I) and R³ is independently —CH₃. In embodiments, the compound hasformula (I) and R³ is independently —CF₃. In embodiments, the compoundhas formula (I) and R³ is independently —OH. In embodiments, thecompound has formula (I) and R³ is independently substituted4-methoxybenzyl. In embodiments, the compound has formula (I) and R³ isindependently substituted 4-methoxyphenyl. In embodiments, the compoundhas formula (I) and R³ is independently substituted pyridyl. Inembodiments, the compound has formula (I) and R³ is independentlyunsubstituted 4-methoxybenzyl. In embodiments, the compound has formula(I) and R³ is independently unsubstituted 4-methoxyphenyl. Inembodiments, the compound has formula (I) and R³ is independentlyunsubstituted pyridyl. In embodiments, the compound has formula (I) andR³ is substituted or unsubstituted alkyl. In embodiments, the compoundhas formula (I) and R³ is substituted or unsubstituted heteroalkyl. Inembodiments, the compound has formula (I) and R³ is substituted orunsubstituted cycloalkyl. In embodiments, the compound has formula (I)and R³ is substituted or unsubstituted heterocycloalkyl. In embodiments,the compound has formula (I) and R³ is substituted or unsubstitutedaryl. In embodiments, the compound has formula (I) and R³ is substitutedor unsubstituted heteroaryl. In embodiments, the compound has formula(I) and R³ is unsubstituted alkyl. In embodiments, the compound hasformula (I) and R³ is unsubstituted heteroalkyl. In embodiments, thecompound has formula (I) and R³ is unsubstituted cycloalkyl. Inembodiments, the compound has formula (I) and R³ is unsubstitutedheterocycloalkyl. In embodiments, the compound has formula (I) and R³ isunsubstituted aryl. In embodiments, the compound has formula (I) and R³is unsubstituted heteroaryl. In embodiments, the compound has formula(I) and R³ is substituted or unsubstituted C₁-C₈ alkyl. In embodiments,the compound has formula (I) and R³ is substituted or unsubstitutedC₁-C₆ alkyl. In embodiments, the compound has formula (I) and R³ issubstituted or unsubstituted C₁-C₄ alkyl. In embodiments, the compoundhas formula (I) and R³ is substituted or unsubstituted C₁-C₂ alkyl. Inembodiments, the compound has formula (I) and R³ is substituted orunsubstituted methyl. In embodiments, the compound has formula (I) andR³ is unsubstituted methyl. In embodiments, the compound has formula (I)and R³ is substituted or unsubstituted 2 to 8 membered heteroalkyl. Inembodiments, the compound has formula (I) and R³ is substituted orunsubstituted 2 to 6 membered heteroalkyl. In embodiments, the compoundhas formula (I) and R³ is substituted or unsubstituted 2 to 4 memberedheteroalkyl. In embodiments, the compound has formula (I) and R³ issubstituted or unsubstituted 2 to 3 membered heteroalkyl. Inembodiments, the compound has formula (I) and R³ is substituted orunsubstituted C₃-C₇ cycloalkyl. In embodiments, the compound has formula(I) and R³ is substituted or unsubstituted 3 to 7 memberedheterocycloalkyl. In embodiments, the compound has formula (I) and R³ issubstituted or unsubstituted 5 to 7 membered heterocycloalkyl. Inembodiments, the compound has formula (I) and R³ is substituted orunsubstituted 5 to 6 membered heterocycloalkyl. In embodiments, thecompound has formula (I) and R³ is substituted or unsubstituted C₆-C₁₀aryl. In embodiments, the compound has formula (I) and R³ is substitutedor unsubstituted phenyl. In embodiments, the compound has formula (I)and R³ is substituted or unsubstituted 5 to 9 membered heteroaryl. Inembodiments, the compound has formula (I) and R³ is substituted orunsubstituted 5 to 6 membered heteroaryl. In embodiments, the compoundhas formula (I) and R³ is substituted or unsubstituted 5 memberedheteroaryl. In embodiments, the compound has formula (I) and R³ issubstituted or unsubstituted 6 membered heteroaryl. In embodiments, thecompound has formula (I) and R³ is substituted or unsubstituted fusedring heteroaryl. In embodiments, the compound has formula (I) and R³ issubstituted or unsubstituted 9 membered heteroaryl. In embodiments, thecompound has formula (I) and R³ is substituted or unsubstituted 10membered heteroaryl. In embodiments, the compound has formula (I) and R³is —OCH₃. In embodiments, the compound has formula (I) and R³ is—CH₂CH₂OH. In embodiments, the compound has formula (I) and R³ is—CH₂CH₃.

In embodiments, the compound has formula (I) and R² is independentlyhydrogen. In embodiments, the compound has formula (I) and R² isindependently halogen. In embodiments, the compound has formula (I) andR² is independently —CH₃. In embodiments, the compound has formula (I)and R² is independently —CF₃. In embodiments, the compound has formula(I) and R² is independently —OH. In embodiments, the compound hasformula (I) and R² is independently substituted 4-methoxybenzyl. Inembodiments, the compound has formula (I) and R² is independentlysubstituted 4-methoxyphenyl. In embodiments, the compound has formula(I) and R² is independently substituted pyridyl. In embodiments, thecompound has formula (I) and R² is independently unsubstituted4-methoxybenzyl. In embodiments, the compound has formula (I) and R² isindependently unsubstituted 4-methoxyphenyl. In embodiments, thecompound has formula (I) and R² is independently unsubstituted pyridyl.In embodiments, the compound has formula (I) and R² is substituted orunsubstituted alkyl. In embodiments, the compound has formula (I) and R²is substituted or unsubstituted heteroalkyl. In embodiments, thecompound has formula (I) and R² is substituted or unsubstitutedcycloalkyl. In embodiments, the compound has formula (I) and R² issubstituted or unsubstituted heterocycloalkyl. In embodiments, thecompound has formula (I) and R² is substituted or unsubstituted aryl. Inembodiments, the compound has formula (I) and R² is substituted orunsubstituted heteroaryl. In embodiments, the compound has formula (I)and R² is unsubstituted alkyl. In embodiments, the compound has formula(I) and R² is unsubstituted heteroalkyl. In embodiments, the compoundhas formula (I) and R² is unsubstituted cycloalkyl. In embodiments, thecompound has formula (I) and R² is unsubstituted heterocycloalkyl. Inembodiments, the compound has formula (I) and R² is unsubstituted aryl.In embodiments, the compound has formula (I) and R² is unsubstitutedheteroaryl. In embodiments, the compound has formula (I) and R² issubstituted or unsubstituted C₁-C₈ alkyl. In embodiments, the compoundhas formula (I) and R² is substituted or unsubstituted C₁-C₆ alkyl. Inembodiments, the compound has formula (I) and R² is substituted orunsubstituted C₁-C₄ alkyl. In embodiments, the compound has formula (I)and R² is substituted or unsubstituted C₁-C₂ alkyl. In embodiments, thecompound has formula (I) and R² is substituted or unsubstituted methyl.In embodiments, the compound has formula (I) and R² is unsubstitutedmethyl. In embodiments, the compound has formula (I) and R² issubstituted or unsubstituted 2 to 8 membered heteroalkyl. Inembodiments, the compound has formula (I) and R² is substituted orunsubstituted 2 to 6 membered heteroalkyl. In embodiments, the compoundhas formula (I) and R² is substituted or unsubstituted 2 to 4 memberedheteroalkyl. In embodiments, the compound has formula (I) and R² issubstituted or unsubstituted 2 to 3 membered heteroalkyl. Inembodiments, the compound has formula (I) and R² is substituted orunsubstituted C₃-C₇ cycloalkyl. In embodiments, the compound has formula(I) and R² is substituted or unsubstituted 3 to 7 memberedheterocycloalkyl. In embodiments, the compound has formula (I) and R² issubstituted or unsubstituted 5 to 7 membered heterocycloalkyl. Inembodiments, the compound has formula (I) and R² is substituted orunsubstituted 5 to 6 membered heterocycloalkyl. In embodiments, thecompound has formula (I) and R² is substituted or unsubstituted C₆-C₁₀aryl. In embodiments, the compound has formula (I) and R² is substitutedor unsubstituted phenyl. In embodiments, the compound has formula (I)and R² is substituted or unsubstituted 5 to 9 membered heteroaryl. Inembodiments, the compound has formula (I) and R² is substituted orunsubstituted 5 to 6 membered heteroaryl. In embodiments, the compoundhas formula (I) and R² is substituted or unsubstituted 5 memberedheteroaryl. In embodiments, the compound has formula (I) and R² issubstituted or unsubstituted 6 membered heteroaryl. In embodiments, thecompound has formula (I) and R² is substituted or unsubstituted fusedring heteroaryl. In embodiments, the compound has formula (I) and R² issubstituted or unsubstituted 9 membered heteroaryl. In embodiments, thecompound has formula (I) and R² is substituted or unsubstituted 10membered heteroaryl. In embodiments, the compound has formula (I) and R²is —OCH₃. In embodiments, the compound has formula (I) and R² is—CH₂CH₂OH. In embodiments, the compound has formula (I) and R² is—CH₂CH₃.

In embodiments, the compound has formula (I) and R¹ and R² are joined toform a substituted or unsubstituted heterocycloalkyl or a substituted orunsubstituted heteroaryl. In embodiments, the compound has formula (I)and R¹ and R² are joined to form a substituted heterocycloalkyl. Inembodiments, the compound has formula (I) and R¹ and R² are joined toform an unsubstituted heterocycloalkyl. In embodiments, the compound hasformula (I) and R¹ and R² are joined to form a substituted heteroaryl.In embodiments, the compound has formula (I) and R¹ and R² are joined toform an unsubstituted heteroaryl. In embodiments, the compound hasformula (I) and R¹ and R² are joined to form a substituted orunsubstituted 3 to 7 membered heterocycloalkyl. In embodiments, thecompound has formula (I) and R¹ and R² are joined to form a substitutedor unsubstituted 5 to 7 membered heterocycloalkyl. In embodiments, thecompound has formula (I) and R¹ and R² are joined to form a substitutedor unsubstituted 5 to 6 membered heterocycloalkyl. In embodiments, thecompound has formula (I) and R¹ and R² are joined to form a substitutedor unsubstituted 5 to 9 membered heteroaryl. In embodiments, thecompound has formula (I) and R¹ and R² are joined to form a substitutedor unsubstituted 5 to 6 membered heteroaryl. In embodiments, thecompound has formula (I) and R¹ and R² are joined to form a substitutedor unsubstituted 5 membered heteroaryl. In embodiments, the compound hasformula (I) and R¹ and R² are joined to form a substituted orunsubstituted 6 membered heteroaryl. In embodiments, the compound hasformula (I) and R¹ and R² are joined to form a substituted orunsubstituted fused ring heteroaryl. In embodiments, the compound hasformula (I) and R¹ and R² are joined to form a substituted orunsubstituted 9 membered heteroaryl. In embodiments, the compound hasformula (I) and R¹ and R² are joined to form a substituted orunsubstituted 10 membered heteroaryl. In embodiments, the compound hasformula (I) and R¹ and R² are joined to form a substituted piperidinyl.In embodiments, the compound has formula (I) and R¹ and R² are joined toform an unsubstituted piperidinyl.

In embodiments, the compound has formula (I) and R² is L²-R⁴. Inembodiments, the compound has formula (I) and L² is unsubstituted C₁-C₁₀alkylene. In embodiments, the compound has formula (I) and L² isunsubstituted C₁-C₈ alkylene. In embodiments, the compound has formula(I) and L² is unsubstituted C₁-C₆ alkylene. In embodiments, the compoundhas formula (I) and L² is unsubstituted C₁-C₄ alkylene. In embodiments,the compound has formula (I) and L² is unsubstituted C₁-C₃ alkylene. Inembodiments, the compound has formula (I) and L² is unsubstituted C₁-C₂alkylene. In embodiments, the compound has formula (I) and L² isunsubstituted branched C₁-C₄ alkylene. In embodiments, the compound hasformula (I) and L² is unsubstituted branched C₁-C₃ alkylene. Inembodiments, the compound has formula (I) and L² is unsubstitutedmethylene. In embodiments, the compound has formula (I) and L² isunsubstituted ethylene. In embodiments, the compound has formula (I) andL² is a bond. In embodiments, the compound has formula (I) and R⁴ isindependently hydrogen. In embodiments, the compound has formula (I) andR⁴ is independently halogen. In embodiments, the compound has formula(I) and R⁴ is independently —CH₃. In embodiments, the compound hasformula (I) and R⁴ is independently —CF₃. In embodiments, the compoundhas formula (I) and R⁴ is independently —OH. In embodiments, thecompound has formula (I) and R⁴ is independently substituted4-methoxybenzyl. In embodiments, the compound has formula (I) and R⁴ isindependently substituted 4-methoxyphenyl. In embodiments, the compoundhas formula (I) and R⁴ is independently substituted pyridyl. Inembodiments, the compound has formula (I) and R⁴ is independentlyunsubstituted 4-methoxybenzyl. In embodiments, the compound has formula(I) and R⁴ is independently unsubstituted 4-methoxyphenyl. Inembodiments, the compound has formula (I) and R⁴ is independentlyunsubstituted pyridyl. In embodiments, the compound has formula (I) andR⁴ is substituted or unsubstituted alkyl. In embodiments, the compoundhas formula (I) and R⁴ is substituted or unsubstituted heteroalkyl. Inembodiments, the compound has formula (I) and R⁴ is substituted orunsubstituted cycloalkyl. In embodiments, the compound has formula (I)and R⁴ is substituted or unsubstituted heterocycloalkyl. In embodiments,the compound has formula (I) and R⁴ is substituted or unsubstitutedaryl. In embodiments, the compound has formula (I) and R⁴ is substitutedor unsubstituted heteroaryl. In embodiments, the compound has formula(I) and R⁴ is unsubstituted alkyl. In embodiments, the compound hasformula (I) and R⁴ is unsubstituted heteroalkyl. In embodiments, thecompound has formula (I) and R⁴ is unsubstituted cycloalkyl. Inembodiments, the compound has formula (I) and R⁴ is unsubstitutedheterocycloalkyl. In embodiments, the compound has formula (I) and R⁴ isunsubstituted aryl. In embodiments, the compound has formula (I) and R⁴is unsubstituted heteroaryl. In embodiments, the compound has formula(I) and R⁴ is substituted or unsubstituted C₁-C₈ alkyl. In embodiments,the compound has formula (I) and R⁴ is substituted or unsubstitutedC₁-C₆ alkyl. In embodiments, the compound has formula (I) and R⁴ issubstituted or unsubstituted C₁-C₄ alkyl. In embodiments, the compoundhas formula (I) and R⁴ is substituted or unsubstituted C₁-C₂ alkyl. Inembodiments, the compound has formula (I) and R⁴ is substituted orunsubstituted methyl. In embodiments, the compound has formula (I) andR⁴ is unsubstituted methyl. In embodiments, the compound has formula (I)and R⁴ is substituted or unsubstituted 2 to 8 membered heteroalkyl. Inembodiments, the compound has formula (I) and R⁴ is substituted orunsubstituted 2 to 6 membered heteroalkyl. In embodiments, the compoundhas formula (I) and R⁴ is substituted or unsubstituted 2 to 4 memberedheteroalkyl. In embodiments, the compound has formula (I) and R⁴ issubstituted or unsubstituted 2 to 3 membered heteroalkyl. Inembodiments, the compound has formula (I) and R⁴ is substituted orunsubstituted C₃-C₇ cycloalkyl. In embodiments, the compound has formula(I) and R⁴ is substituted or unsubstituted 3 to 7 memberedheterocycloalkyl. In embodiments, the compound has formula (I) and R⁴ issubstituted or unsubstituted 5 to 7 membered heterocycloalkyl. Inembodiments, the compound has formula (I) and R⁴ is substituted orunsubstituted 5 to 6 membered heterocycloalkyl. In embodiments, thecompound has formula (I) and R⁴ is substituted or unsubstituted C₆-C₁₀aryl. In embodiments, the compound has formula (I) and R⁴ is substitutedor unsubstituted phenyl. In embodiments, the compound has formula (I)and R⁴ is substituted or unsubstituted 5 to 9 membered heteroaryl. Inembodiments, the compound has formula (I) and R⁴ is substituted orunsubstituted 5 to 6 membered heteroaryl. In embodiments, the compoundhas formula (I) and R⁴ is substituted or unsubstituted 5 memberedheteroaryl. In embodiments, the compound has formula (I) and R⁴ issubstituted or unsubstituted 6 membered heteroaryl. In embodiments, thecompound has formula (I) and R⁴ is substituted or unsubstituted fusedring heteroaryl. In embodiments, the compound has formula (I) and R⁴ issubstituted or unsubstituted 9 membered heteroaryl. In embodiments, thecompound has formula (I) and R⁴ is substituted or unsubstituted 10membered heteroaryl. In embodiments, the compound has formula (I) and R⁴is —OCH₃. In embodiments, the compound has formula (I) and R⁴ is—CH₂CH₂OH. In embodiments, the compound has formula (I) and R⁴ is—CH₂CH₃.

In embodiments, the compound has formula (Ia) and L⁰, L¹, L², R¹, R²,R³, and R⁴ of any one of the embodiments for the compound of formula (I)described herein above.

In embodiments, the compound has formula (II) and L⁰, L¹, R¹, and R³ ofany one of the embodiments for the compound of formula (I) describedherein above.

In embodiments, the compound has formula (IIa) and L⁰, L¹, R¹, and R³ ofany one of the embodiments for the compound of formula (I) describedherein above.

In embodiments, the compound has formula (III) and L⁰, L¹, R¹, and R³ ofany one of the embodiments for the compound of formula (I) describedherein above.

In embodiments, the compound has formula (IIIa) and L⁰, L¹, R¹, and R³of any one of the embodiments for the compound of formula (I) describedherein above.

In embodiments, the compound has formula (IV) and L⁰, L¹, L², R¹, R²,R³, and R⁴ of any one of the embodiments for the compound of formula (I)described herein above.

In embodiments, the compound has formula (IVa) and L⁰, L¹, L², R¹, R²,R³, and R⁴ of any one of the embodiments for the compound of formula (I)described herein above.

In embodiments, the compound is selected from the group consisting of

In some embodiments, the compound is selected from the group consistingof JD202, JD203, JD204, JD205, JD206, JD207, JD208, JD209, and JD2010(including from the neutral compound, protonated cationic compound, andpharmaceutically acceptable salt of the compound). In embodiments, thecompound is not JD201 (i.e. DMAPT). In embodiments, the compound is notthe neutral form of JD201. In embodiments, the compound is not aprotonated cationic form of JD201. In embodiments, the compound is not afumarate salt of JD201. In embodiments, the compound is not apharmaceutically acceptable salt of JD201.

In embodiments, the compound is not a compound of formula (I) or (Ia)wherein L⁰ is unsubstituted methylene and R¹ and R² are independentlyhydrogen or unsubstituted methyl or ethyl. In embodiments, the compoundis not a compound of formula (I) or (Ia) wherein L⁰ is unsubstitutedmethylene and R¹ and R² are unsubstituted ethyl. In embodiments, thecompound is not a compound of formula (I) or (Ia) wherein L⁰ isunsubstituted methylene and R¹ and R² are independently hydrogen orunsubstituted C₁-C₃ alkyl. In embodiments, the compound is not acompound of formula (I) or (Ia) wherein L⁰ is unsubstituted methyleneand R¹ and R² are independently hydrogen or unsubstituted linear C₁-C₃alkyl. In embodiments, the compound is not a compound of formula (I) or(Ia) wherein L⁰ is unsubstituted methylene and R¹ and R² areindependently hydrogen or unsubstituted C₁-C₄ alkyl. In embodiments, thecompound is not a compound of formula (I) or (Ia) wherein L⁰ isunsubstituted methylene and R¹ and R² are independently hydrogen orunsubstituted linear C₁-C₄ alkyl. In embodiments, the compound is not acompound of formula (I) or (Ia) wherein L⁰ is unsubstituted methyleneand R¹ and R² are independently hydrogen or halogen substituted methylor unsubstituted methyl. In embodiments, the compound is not a compoundof formula (I) or (Ia) wherein L⁰ is unsubstituted methylene and R¹ andR² are independently hydrogen or halogen substituted C₁-C₂ alkyl orunsubstituted C₁-C₂ alkyl. In embodiments, the compound is not acompound of formula (I) or (Ia) wherein L⁰ is unsubstituted methyleneand R¹ and R² are independently hydrogen or halogen substituted C₁-C₃alkyl or unsubstituted C₁-C₃ alkyl. In embodiments, the compound is nota compound of formula (I) or (Ia) wherein L⁰ is unsubstituted methyleneand R¹ and R² are independently hydrogen or unsubstituted methyl ormethyl substituted with —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH,—NO₂, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, or —NHOH. In embodiments,the compound is not a compound of formula (I) or (Ia) wherein L⁰ isunsubstituted methylene and R¹ and R² are independently hydrogen,unsubstituted C₁-C₂ alkyl or C₁-C₂ alkyl substituted with —CF₃, —CCl₃,—CN, —S(O)CH₃, —NH₂, —COOH, —CONH₂, —NO₂, —SH, —SO₂Cl, —SO₃H, —SO₄H,—SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H,—NHC(O)OH, or —NHOH.

In embodiments, the compound is not a compound of formula (I) or (Ia)wherein L⁰ is a bond and R¹ and R² are independently hydrogen orunsubstituted methyl or ethyl. In embodiments, the compound is not acompound of formula (I) or (Ia) wherein L⁰ is a bond and R¹ and R² areunsubstituted ethyl. In embodiments, the compound is not a compound offormula (I) w or (Ia) herein L⁰ is a bond and R¹ and R² areindependently hydrogen or unsubstituted C₁-C₃ alkyl. In embodiments, thecompound is not a compound of formula (I) or (Ia) wherein L⁰ is a bondand R¹ and R² are independently hydrogen or unsubstituted linear C₁-C₃alkyl. In embodiments, the compound is not a compound of formula (I) wor (Ia) herein L⁰ is a bond and R¹ and R² are independently hydrogen orunsubstituted C₁-C₄ alkyl. In embodiments, the compound is not acompound of formula (I) or (Ia) wherein L⁰ is a bond and R¹ and R² areindependently hydrogen or unsubstituted linear C₁-C₄ alkyl. Inembodiments, the compound is not a compound of formula (I) or (Ia)wherein L⁰ is a bond and R¹ and R² are independently hydrogen or halogensubstituted methyl or unsubstituted methyl. In embodiments, the compoundis not a compound of formula (I) w or (Ia) herein L⁰ is a bond and R¹and R² are independently hydrogen or halogen substituted C₁-C₂ alkyl orunsubstituted C₁-C₂ alkyl. In embodiments, the compound is not acompound of formula (I) or (Ia) wherein L⁰ is a bond and R¹ and R² areindependently hydrogen or halogen substituted C₁-C₃ alkyl orunsubstituted C₁-C₃ alkyl. In embodiments, the compound is not acompound of formula (I) or (Ia) wherein L⁰ is a bond and R¹ and R² areindependently hydrogen or unsubstituted methyl or methyl substitutedwith —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —SH,—SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, or —NHOH.

In embodiments, the compound is not a compound of formula (I) or (Ia)wherein L⁰ is unsubstituted C₁-C₂ alkylene and R¹ and R² areindependently hydrogen or unsubstituted methyl or ethyl. In embodiments,the compound is not a compound of formula (I) or (Ia) wherein L⁰ isindependently hydrogen or unsubstituted C₁-C₂ alkylene and R¹ and R² areunsubstituted ethyl. In embodiments, the compound is not a compound offormula (I) or (Ia) wherein L⁰ is unsubstituted C₁-C₂ alkylene and R¹and R² are independently hydrogen or unsubstituted C₁-C₃ alkyl. Inembodiments, the compound is not a compound of formula (I) w or (Ia)herein L⁰ is unsubstituted C₁-C₂ alkylene and R¹ and R² areindependently hydrogen or unsubstituted linear C₁-C₃ alkyl. Inembodiments, the compound is not a compound of formula (I) or (Ia)wherein L⁰ is unsubstituted C₁-C₂ alkylene and R¹ and R² areindependently hydrogen or unsubstituted C₁-C₄ alkyl. In embodiments, thecompound is not a compound of formula (I) or (Ia) herein L⁰ isunsubstituted C₁-C₂ alkylene and R¹ and R² are independently hydrogen orunsubstituted linear C₁-C₄ alkyl. In embodiments, the compound is not acompound of formula (I) or (Ia) wherein L⁰ is unsubstituted C₁-C₂alkylene and R¹ and R² are independently hydrogen or halogen substitutedmethyl or unsubstituted methyl. In embodiments, the compound is not acompound of formula (I) or (Ia) wherein L⁰ is unsubstituted C₁-C₂alkylene and R¹ and R² are independently hydrogen or halogen substitutedC₁-C₂ alkyl or unsubstituted C₁-C₂ alkyl. In embodiments, the compoundis not a compound of formula (I) or (Ia) wherein L⁰ is unsubstitutedC₁-C₂ alkylene and R¹ and R² are independently hydrogen or halogensubstituted C₁-C₃ alkyl or unsubstituted C₁-C₃ alkyl. In embodiments,the compound is not a compound of formula (I) or (Ia) wherein L⁰ isunsubstituted C₁-C₂ alkylene and R¹ and R² are independently hydrogen orunsubstituted methyl or methyl substituted with —CF₃, —CCl₃, —CN,—S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —SH, —SO₂Cl, —SO₃H, —SO₄H,—SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂, NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H,—NHC(O)OH, or —NHOH.

III. Pharmaceutical Compositions

In another aspect is provided a pharmaceutical composition including apharmaceutically acceptable excipient and a compound, or apharmaceutically acceptable salt thereof, as described herein (includingembodiments).

In embodiments of the pharmaceutical compositions, the compound (e.g. asdescribed herein, including embodiments), or pharmaceutically acceptablesalt thereof, is included in a therapeutically effective amount. Inembodiments of the pharmaceutical compositions, the pharmaceuticalcomposition includes a second agent (e.g. therapeutic agent). Inembodiments of the pharmaceutical compositions, the pharmaceuticalcomposition includes a second agent (e.g. therapeutic agent) in atherapeutically effective amount. In embodiments, the second agent is anagent for treating cancer (e.g. leukemia, lung cancer, epidermoidcarcinoma (i.e. squamous-cell carcinoma), fibrosarcoma, liver cancer(e.g. hepatocellular carcinoma), prostate cancer, kidney cancer (e.g.renal cell carcinoma or urothelial cell carcinoma), lymphoma, breastcancer, urinary bladder cancer, prostate cancer, or therapy resistantcancer). In embodiments, the second agent is an anti-cancer agent. Inembodiments, the second agent is a chemotherapeutic. In embodiments, thesecond agent is a hormonal therapeutic agent. In embodiments, the secondagent is a platinum-based compound. In embodiments, the second agent istamoxifen. In embodiments, the second agent is trastuzumab. Inembodiments, the second agent is cisplatin. In embodiments, the secondagent is carboplatin. In embodiments, the second agent is an agent fortreating leukemia. In embodiments, the second agent is an agent fortreating breast cancer. In embodiments, the second agent is an agent fortreating lung cancer. In embodiments, the second agent is an agent fortreating non-small cell lung cancer. In embodiments, the second agent isan agent for treating a therapy resistant cancer. In embodiments, thesecond agent is an agent for reducing NFκB phosphorylation. Inembodiments, the second agent is an agent for inhibiting a pathwayactivated by NFκB phosphorylation. In embodiments, the second agent isan agent for reducing NFκB activity. In embodiments, the second agent isan agent for treating tamoxifen resistant cancer. In embodiments, thesecond agent is an agent for treating cisplatin resistant cancer. Inembodiments, the second agent is an agent for treating carboplatinresistant cancer. In embodiments, the second agent is an agent fortreating trastuzumab resistant cancer. In embodiments, the second agentis an agent for treating hormonal therapy resistant cancer. Inembodiments, the second agent is an agent for treating platinum-basedcompound resistant cancer. In embodiments, the second agent is an agentfor treating a hyperproliferative disease (e.g. cancer or anon-malignant hyperproliferative disease). In embodiments, the secondagent is an agent for treating a hamartomatous lesion. In embodiments,the second agent is an agent for treating angiomyolipoma. Inembodiments, the second agent is an agent for treatinglymphangioleiomyomatosis. In embodiments, the second agent is an agentfor treating tuberous sclerosis complex. In embodiments, the secondagent is an agent for treating a hamartia. In embodiments, the secondagent is an agent for treating a hamartoma. In embodiments, the secondagent is an agent for increasing the activity of a pathway includingTSC1. In embodiments, the second agent is an agent for increasing TSC1activity. In embodiments, the second agent is an agent for increasingthe activity of a pathway including TSC2. In embodiments, the secondagent is an agent for increasing TSC2 activity. In embodiments, thesecond agent is an agent for decreasing the activity of a pathwayincluding mTOR. In embodiments, the second agent is an agent fordecreasing mTOR activity. In embodiments, the second agent is an agentfor decreasing the activity of a pathway including mTORC1. Inembodiments, the second agent is an agent for decreasing mTORC1activity.

In some embodiments, the compound is a compound described in an example,a table, a figure, or a claim. In some embodiments, the compound is acompound described in the Compounds section above.

IV. Methods of Treatment

In another aspect is provided a method of treating cancer in a patientin need of the treatment, the method including administering atherapeutically effective amount of a compound, or a pharmaceuticallyacceptable salt thereof, as described herein (including embodiments) tothe patient.

In embodiments, the cancer is breast cancer. In embodiments, the canceris a therapy resistant breast cancer. In embodiments, the cancer is ahormonal therapy resistant breast cancer. In embodiments, the cancer istamoxifen resistant breast cancer. In embodiments, the cancer istrastuzumab (i.e. Herceptin™) resistant breast cancer. In embodiments,the cancer is breast cancer resistant to an aromatase inhibitor. Inembodiments, the cancer is fulvestrant resistant breast cancer. Inembodiments, the cancer is exemestane resistant breast cancer. Inembodiments, the cancer is anastrozole resistant breast cancer. Inembodiments, the cancer is aminoglutethimide resistant breast cancer. Inembodiments, the cancer is testolactone resistant breast cancer. Inembodiments, the cancer is letrozole resistant breast cancer. Inembodiments, the cancer is vorozole resistant breast cancer. Inembodiments, the cancer is formestane resistant breast cancer. Inembodiments, the cancer is fadrozole resistant breast cancer. Inembodiments, the cancer is metastatic breast cancer. In embodiments, thecancer is ER positive breast cancer. In embodiments, the cancer is ERnegative breast cancer. In embodiments, the cancer is breast cancerresistant to an anti-cancer agent. In embodiments, the cancer is breastcancer expressing a high level of HER2 (e.g. relative to a control suchas a non-cancerous sample).

In embodiments, the cancer is lung cancer. In embodiments, the cancer isnon-small cell lung cancer (NSCLC). In embodiments, the cancer is atherapy resistant NSCLC. In embodiments, the cancer is oxaliplatinresistant NSCLC. In embodiments, the cancer is cisplatin resistantNSCLC. In embodiments, the cancer is NSCLC resistant to a platinum-basedcompound. In embodiments, the cancer is carboplatin resistant NSCLC. Inembodiments, the cancer is lung cancer resistant to a platinum-basedcompound. In embodiments, the cancer is metastatic lung cancer. Inembodiments, the cancer is metastatic NSCLC. In embodiments, the canceris lung cancer resistant to an anti-cancer agent. In embodiments, thecancer is NSCLC resistant to an anti-cancer agent. In embodiments, thecancer is resistant to a taxane. In embodiments, the cancer is resistantto a hormonal therapy. In embodiments, the cancer is resistant to aplatinum-based compound. In embodiments, the cancer is resistant todocetaxel.

In embodiments, the cancer is urinary bladder cancer. In embodiments,the cancer is a therapy resistant urinary bladder cancer. Inembodiments, the cancer is prostate cancer. In embodiments, the canceris a therapy resistant prostate cancer. In embodiments, the cancer isleukemia. In embodiments, the cancer is a therapy resistant leukemia. Inembodiments, the cancer is lymphoma. In embodiments, the cancer is atherapy resistant lymphoma. In embodiments, the cancer is epidermoidcarcinoma (i.e. squamous-cell carcinoma). In embodiments, the cancer isa therapy resistant epidermoid carcinoma (i.e. squamous-cell carcinoma).In embodiments, the cancer is fibrosarcoma. In embodiments, the canceris a therapy resistant fibrosarcoma. In embodiments, the cancer is livercancer (e.g. hepatocellular carcinoma). In embodiments, the cancer is atherapy resistant liver cancer (e.g. hepatocellular carcinoma). Inembodiments, the cancer is kidney cancer (e.g. renal cell carcinoma orurothelial cell carcinoma). In embodiments, the cancer is a therapyresistant kidney cancer (e.g. renal cell carcinoma or urothelial cellcarcinoma). In embodiments, the cancer is a hematopoietic cell cancer.In embodiments, the cancer is a therapy resistant hematopoietic cellcancer. In embodiments, the cancer is metastatic cancer. In embodiments,the cancer is resistant to an anti-cancer agent.

In another aspect is provided a method of treating a non-malignanthyperproliferative disease in a patient in need of the treatment, themethod including administering a therapeutically effective amount of acompound, or a pharmaceutically acceptable salt thereof, as describedherein (including embodiments) to the patient.

In embodiments, the non-malignant hyperproliferative disease is ahamartomatous lesion. In embodiments, the non-malignanthyperproliferative disease is angiomyolipoma. In embodiments, thenon-malignant hyperproliferative disease is lymphangioleiomyomatosis. Inembodiments, the non-malignant hyperproliferative disease is tuberoussclerosis complex. In embodiments, the non-malignant hyperproliferativedisease is a hamartia. In embodiments, the non-malignanthyperproliferative disease is a hamartoma.

In embodiments of the methods of treating a disease (e.g. cancer or anon-malignant hyperproliferative disease), the compound, orpharmaceutically acceptable salt thereof, is provided as apharmaceutical composition (as described herein, including embodiments).

In embodiments, the compound, or a pharmaceutically acceptable saltthereof, is co-administered with a second agent (e.g. therapeuticagent). In embodiments, the compound, or a pharmaceutically acceptablesalt thereof, is co-administered with a second agent (e.g. therapeuticagent), which is administered in a therapeutically effective amount. Inembodiments, the second agent is an agent for treating cancer (e.g.leukemia, lung cancer, epidermoid carcinoma (i.e. squamous-cellcarcinoma), fibrosarcoma, liver cancer (e.g. hepatocellular carcinoma),prostate cancer, kidney cancer (e.g. renal cell carcinoma or urothelialcell carcinoma), lymphoma, breast cancer, urinary bladder cancer,prostate cancer, or therapy resistant cancer). In embodiments, thesecond agent is an anti-cancer agent. In embodiments, the second agentis a chemotherapeutic. In embodiments, the second agent is a hormonaltherapeutic agent. In embodiments, the second agent is a platinum-basedcompound. In embodiments, the second agent is tamoxifen. In embodiments,the second agent is trastuzumab. In embodiments, the second agent iscisplatin. In embodiments, the second agent is carboplatin. Inembodiments, the second agent is an agent for treating leukemia. Inembodiments, the second agent is an agent for treating breast cancer. Inembodiments, the second agent is an agent for treating lung cancer. Inembodiments, the second agent is an agent for treating non-small celllung cancer. In embodiments, the second agent is an agent for treating atherapy resistant cancer (e.g. a therapy resistant leukemia, lungcancer, epidermoid carcinoma (i.e. squamous-cell carcinoma),fibrosarcoma, liver cancer (e.g. hepatocellular carcinoma), prostatecancer, kidney cancer (e.g. renal cell carcinoma or urothelial cellcarcinoma), lymphoma, breast cancer, urinary bladder cancer, or prostatecancer). In embodiments, the second agent is an agent for reducing NFκBphosphorylation. In embodiments, the second agent is an agent forinhibiting a pathway activated by NFκB phosphorylation. In embodiments,the second agent is an agent for reducing NFκB activity. In embodiments,the second agent is an agent for treating tamoxifen resistant cancer. Inembodiments, the second agent is an agent for treating cisplatinresistant cancer. In embodiments, the second agent is an agent fortreating carboplatin resistant cancer. In embodiments, the second agentis an agent for treating trastuzumab resistant cancer. In embodiments,the second agent is an agent for treating hormonal therapy resistantcancer. In embodiments, the second agent is an agent for treatingplatinum-based compound resistant cancer. In embodiments, the secondagent is an agent for treating a hyperproliferative disease (e.g. canceror a non-malignant hyperproliferative disease). In embodiments, thesecond agent is an agent for treating a hamartomatous lesion. Inembodiments, the second agent is an agent for treating angiomyolipoma.In embodiments, the second agent is an agent for treatinglymphangioleiomyomatosis. In embodiments, the second agent is an agentfor treating tuberous sclerosis complex. In embodiments, the secondagent is an agent for treating a hamartia. In embodiments, the secondagent is an agent for treating a hamartoma. In embodiments, the secondagent is an agent for increasing the activity of a pathway includingTSC1. In embodiments, the second agent is an agent for increasing TSC1activity. In embodiments, the second agent is an agent for increasingthe activity of a pathway including TSC2. In embodiments, the secondagent is an agent for increasing TSC2 activity. In embodiments, thesecond agent is an agent for decreasing the activity of a pathwayincluding mTOR. In embodiments, the second agent is an agent fordecreasing mTOR activity. In embodiments, the second agent is an agentfor decreasing the activity of a pathway including mTORC1. Inembodiments, the second agent is an agent for decreasing mTORC1activity.

In some embodiments, the compound is a compound described in an example,a table, a figure, or a claim. In some embodiments, the compound is acompound described in the Compounds section above.

V. Methods of Modulating a Target

In another aspect is provided a method of inhibiting cancer cell growthor survival including contacting the cell with an effective amount of acompound, or a pharmaceutically acceptable salt thereof, as describedherein (including embodiments).

In embodiments, the cancer cell is a stem cell. In embodiments, thecancer cell is a progenitor cell. In embodiments, the cancer cell is acancer (e.g. leukemia, lung cancer, epidermoid carcinoma (i.e.squamous-cell carcinoma), fibrosarcoma, liver cancer (e.g.hepatocellular carcinoma), prostate cancer, kidney cancer (e.g. renalcell carcinoma or urothelial cell carcinoma), lymphoma, breast cancer,urinary bladder cancer, prostate cancer, or therapy resistant cancer)stem cell. In embodiments, the cancer cell is a cancer (e.g. leukemia,lung cancer, epidermoid carcinoma (i.e. squamous-cell carcinoma),fibrosarcoma, liver cancer (e.g. hepatocellular carcinoma), prostatecancer, kidney cancer (e.g. renal cell carcinoma or urothelial cellcarcinoma), lymphoma, breast cancer, urinary bladder cancer, prostatecancer, or therapy resistant cancer) progenitor cell. In embodiments,the cancer cell is a lung cancer stem cell. In embodiments, the cancercell is a lung cancer progenitor cell. In embodiments, the cancer cellis a breast cancer stem cell. In embodiments, the cancer cell is abreast cancer progenitor cell. In embodiments, the compound, or apharmaceutically acceptable salt thereof, (as described herein,including embodiments) increases apoptosis in the cancer cell. Inembodiments, the compound, or a pharmaceutically acceptable saltthereof, (as described herein, including embodiments) induces apoptosisin the cancer cell.

In another aspect is provided a method of modulating the level ofactivity of NF-κB in a cell including contacting the cell with aneffective amount of a compound, or a pharmaceutically acceptable saltthereof, as described herein (including embodiments).

In embodiments of the methods of modulating NFκB, modulating isinhibiting the level of activity of NFκB. In embodiments of the methodsof modulating NFκB, modulating is increasing the level of activity ofNFκB. In embodiments of the methods of modulating NFκB, the compound, orpharmaceutically acceptable salt thereof, is provided as apharmaceutical composition (as described herein, including embodiments).In embodiments of the method of modulating NFκB, the method includescontacting NFκB with a compound as described herein (includingembodiments). In embodiments of the method of modulating NFκB, themethod includes contacting a component of a pathway including NFκB witha compound as described herein (including embodiments). In embodimentsof the methods of modulating NFκB, modulating is inhibiting thephosphorylation of NFκB. In embodiments of the methods of modulatingNFκB, modulating is inhibiting a pathway activated by thephosphorylation of NFκB. In embodiments of the methods of modulatingNFκB, modulating is inhibiting a pathway including phosphorylated NFκB.

In another aspect is provided a method of modulating the level ofactivity of TSC1 in a cell including contacting the cell with aneffective amount of a compound, or a pharmaceutically acceptable saltthereof, as described herein (including embodiments).

In embodiments of the methods of modulating TSC1, modulating isinhibiting the level of activity of TSC1. In embodiments of the methodsof modulating TSC1, modulating is increasing the level of activity ofTSC1. In embodiments of the methods of modulating TSC1, the compound, orpharmaceutically acceptable salt thereof, is provided as apharmaceutical composition (as described herein, including embodiments).In embodiments of the method of modulating TSC1, the method includescontacting TSC1 with a compound as described herein (includingembodiments). In embodiments of the method of modulating TSC1, themethod includes modulating the level of activity or amount of acomponent in a pathway including TSC1.

In another aspect is provided a method of modulating the level ofactivity of TSC2 in a cell including contacting the cell with aneffective amount of a compound, or a pharmaceutically acceptable saltthereof, as described herein (including embodiments).

In embodiments of the methods of modulating TSC2, modulating isinhibiting the level of activity of TSC2. In embodiments of the methodsof modulating TSC2, modulating is increasing the level of activity ofTSC2. In embodiments of the methods of modulating TSC2, the compound, orpharmaceutically acceptable salt thereof, is provided as apharmaceutical composition (as described herein, including embodiments).In embodiments of the method of modulating TSC2, the method includescontacting TSC2 with a compound as described herein (includingembodiments). In embodiments of the method of modulating TSC2, themethod includes modulating the level of activity or amount of acomponent in a pathway including TSC2.

In another aspect is provided a method of modulating the level ofactivity of mTOR in a cell including contacting the cell with aneffective amount of a compound, or a pharmaceutically acceptable saltthereof, as described herein (including embodiments).

In embodiments of the methods of modulating mTOR, modulating isinhibiting the level of activity of mTOR. In embodiments of the methodsof modulating mTOR, modulating is increasing the level of activity ofmTOR. In embodiments of the methods of modulating mTOR, the compound, orpharmaceutically acceptable salt thereof, is provided as apharmaceutical composition (as described herein, including embodiments).In embodiments of the method of modulating mTOR, the method includescontacting mTOR with a compound as described herein (includingembodiments). In embodiments of the method of modulating mTOR, themethod includes modulating the level of activity or amount of acomponent in a pathway including mTOR. In embodiments of the methods ofmodulating mTOR, modulating is inhibiting the level of activity ofmTORC1. In embodiments of the methods of modulating mTOR, modulating isincreasing the level of activity of mTORC1. In embodiments of themethods of modulating mTOR, the compound, or pharmaceutically acceptablesalt thereof, is provided as a pharmaceutical composition (as describedherein, including embodiments). In embodiments of the method ofmodulating mTOR, the method includes contacting mTORC1 with a compoundas described herein (including embodiments). In embodiments of themethod of modulating mTOR, the method includes modulating the level ofactivity or amount of a component in a pathway including mTORC1.

In embodiments, the compound, or a pharmaceutically acceptable saltthereof, is co-administered with a second agent (e.g. therapeuticagent). In embodiments, the compound, or a pharmaceutically acceptablesalt thereof, is co-administered with a second agent (e.g. therapeuticagent), which is administered in a therapeutically effective amount. Inembodiments, the second agent is a second agent as described herein,including in the method of treatment section above and includingembodiments.

In some embodiments, the compound is a compound described in an example,a table, a figure, or a claim. In some embodiments, the compound is acompound described in the Compounds section above.

Lung cancer is the leading cause of cancer death in men and womenworldwide. The poor prognosis of advanced non-small cell lung cancer(NSCLC) is due, in part, to emergence of tumor resistance tochemotherapy. Recent data indicate that human tumors, including NSCLC,contain a small subset of cancer stem/progenitor cells (CSC) responsiblefor drug resistance and tumor maintenance. If such minute subsets of CSCdrive tumor formation and drug resistance, therapies targeting the bulktumor mass but not CSC will fail. We now confirm identification ofsubpopulations of chemotherapy-resistant human NSCLC cells withenrichment for CSC biomarkers and exhibiting significant CSC activity.We identified CD133+/ALDH+ tumor stem/progenitor cells from human lungcancer cells in vitro using established Aldefluor assays in combinationwith labeled anti-CD133 antibodies. Estrogen, a known risk factor forlung cancer progression, stimulated a modest increase in the numbers ofCSC. In contrast to control CD133−/ALDH− tumor cell subsets, CSCsubpopulations grew as tumor spheres and maintained self-renewalcapacity in vitro and exhibited a greater tumorigenic capability thannon-CSC subsets in vivo, properties indicative of CSC. Furthermore,resistance of CSC-like cells to cisplatin (a standard chemotherapy forNSCLC treatment) was fully reversed by treatment with parthenolide(PTL), a naturally-occurring sesquiterpene lactone compound with strongantitumor activity in leukemia and prostate cancer, while sparing normalcells. The antitumor effect of PTL may be due to its action as a potentinhibitor of nuclear factor-KB (NF-κB), which is markedly activated bychemotherapy. To target CSC and suppress tumor progression, wesynthesized and tested novel analogs of PTL with improved antitumorproperties and aqueous solubility. PTL analogs inhibit proliferation ofH157 NSCLC cells using both bulk cell preparations andCSC-subpopulations, with effects significantly different from control atP<0.05. Dose-dependent increments of PTL analogs increase apoptosis ofCSC when compared with bulk cells. Moreover, PTL analogs inhibit cellproliferation of H23, A549 and H1975 NSCLC cells with known resistanceto cisplatin (P<0.01). These compounds were able to sensitize cells tocisplatin-induced cytotoxicity (P<0.01) when cells were exposed tosub-optimal concentrations of cisplatin. Using Western blots, we findthat PTL congeners inhibit phosphorylation of the p65 subunit ofphospho-NF-κB and activation of IKKα/β. Thus, targeted inhibition ofNF-κB may reverse tumor drug resistance by interfering with known NF-κBactions to regulate genes involved in proliferation, DNA damageresponse, antiapoptosis and angiogenesis. Further development of PTLanalogs as therapeutics may lead to new strategies to treat NSCLC in theclinic.

Chemical modification of parthenolide to enhance its solubility allowsthe compound to better suppress not only human lung tumor growth invitro but also tumor xenograft growth in vivo (2,10). We have chemicallysynthesized soluble drug-like parthenolide derivatives and tested themin NSCLC cell lines. These novel analogues are able to suppress NSCLCcell proliferation and inhibit activation of NFκB. This inventionprovides a new method to treat NSCLC and possibly other types of cancersthrough inhibition of NFκB signaling and activation of cell deathpathways. Orally available drug-like parthenolide derivatives could beused as new therapeutics for the treatment of lung cancer andpotentially other solid tumors.

An aminoanalog of parthenolide, dimethylaminoparthenolide (DMAPT/LC-1was demonstrated to have 70% oral bioavaliability, plasma concentrationsof 40 micromolar after oral administration and an acceptable toxicologyprofile (3). DMAPT appears to be active against leukemia, lung, urinarybladder and prostate cancer cells, including cancer stem cells (2,3-5).We have synthesized novel derivatives of the parental parthenolide andDMAPT that we find to be very active in inhibiting proliferation ofNSCLC and inducing cancer stem cell death.

We have synthesized parthenolide derivatives that are unique in theirstructures and activity. These novel compounds have shown to be equallyor more effective than DMAPT and more water-soluble than DMAPT (1). Atpresent, there are no drug-like parthenolides used for treatment of lungcancer or other solid tumors. Development of novel therapies fortreatment of lung cancer and other solid tumors are urgently needed. Ofnote, there are currently no targeted therapies for NSCLC, thepredominant form of lung cancer. Identification of a new targetedtherapeutic for lung and other solid tumors would be significant. It isnotable that a large proportion of antitumor therapeutics currently usedin the clinic were developed from natural products and frominvestigation of the use of herbal constituents in traditionalmedicines. Aminoparthenolide analogues with oral bioavailability inducecancer cell death without having detrimental effects on normal cells.This invention is intended to advance novel chemical compounds withclinical-translational potential for the benefit of patients with NSCLCand other solid tumors. Given the large population of patients afflictedwith NSCLC worldwide, the need for a new treatment strategy in theclinic is urgent. The compounds disclosed herein may be effective intreating other types of solid malignancies such as breast and prostatecancer. Compounds described herein may be useful in cancer preventionamong individuals at high risk for cancer (for example, those with asignificant history of tobacco smoking or evidence of premalignant lunglesions). Oral administration of the novel compounds may be used inclinical practice.

We have developed a set of novel analogues of parthenolide which haveboth improved aqueous solubility and antitumor properties. An extensivestructure-activity study was done to investigate parthenolide and itsanalogues. These new compounds are more water soluble than the parentalparthenolide and even though the structure is altered significantly,they retain enhanced activity over the parental compound. We havedeveloped some analogues (such as JD203) with improved water solubilityand strong anticancer activity as compared to other known parthenolidederivatives such as DMAPT (JD201) (1,2). These water-soluble compounds,for example analog JD206, may be conjugated to sugars or peptides thatmay be selectively targeted to cancer cells, particularly cancer stemcells.

Novel parthenolide derivatives for treatment of NSCLC are being testedin a panel of cell line models of lung cancer. In addition, novelparthenolide derivatives are being tested for their toxicity on normalcells such as blood vessels cells (human umbilical vein endothelialcells) and lung cells (human bronquial epithelial cells). Analogues ofthe present invention may be used alone or in combination with standardchemotherapy treatments currently used in the clinic, such as cisplatin,carboplatin and taxanes. Analogues of the present invention may be usedin the treatment or prevention of lung cancer.

We determined that parthenolide analogues prepared in our laboratoryinhibit phosphorylation of NF-κB/p65. Hence, H23 lung tumor cells weretreated with vehicle (CON) or 10 mM parthenolide analogs (JD201, 202,203, 204 and 205). After 60 min, Western blots were done usinganti-phospho-NFκB/p65 antibody (Cell Signaling). Results are shown inFIG. 13.

Lung Cancer has one of the lowest survival outcomes of all cancers withoverall 5-year survival rate for non-small cell lung cancer at 15%. Thepoor prognosis of advanced non-small cell lung cancer (NSCLC) is due, inpart, to emergence of tumor resistance to chemotherapy. Human tumorscontain a small subset of cancer stem cells (CSC) responsible for drugresistance and tumor maintenance. It is important to design effectivetherapies to target CSC. Parthenolide and analogues enhanced sensitivityto chemotherapy in A549 NSCLC cells by blocking NF-κB signaling.Aberrant activation of NF-κB has been proposed as the major cause ofchemoresistance in lung cancer. Parthenolides combined with low-dosechemotherapy may limit cytotoxicity and elicit enhanced antitumorefficacy by blocking NF-κB activation in lung cancer therapy. Subsets ofdrug-resistant CSCs are present in lung tumors and this resistancephenotype associates with NF-kB signaling to block apoptosis. Use ofparthenolide analogues to selectively inhibit NF-κB and reverseantiapoptotic activity may be a new strategy to eradicate lung cancer.Parthenolide analogues inhibit growth of male and female NSCLC cells invitro. NSCLC cells can be sensitized to cisplatin mediated cytotoxicityby parthenolide derivatives. Cancer stem/progenitor cells occur in thebulk of NSCLC cells and have a response to estrogen exposure.ALDH+/CD133+ tumor cell subpopulations form tumor spheres and exhibitself-renewal properties. Parthenolides inhibit phosphorylation of thep65 subunit of NFκB. Parthenolide analogues block activation of NFκB andare active in restoring breast tumor responses to hormonal andTrastuzumab therapies, thereby potentially helping to improve patientoutcomes. Parthenolide analogues are active in suppressing lung cancerstem cells and have the potential to reverse tumor recurrences afterprimary therapy.

VI. Additional Embodiments

1. A compound, or a pharmaceutically acceptable salt thereof, whereinthe compound has a formula selected from the group consisting of:

wherein, L⁰ is independently a bond or an unsubstituted C₁-C₁₀ alkylene;and R¹ and R² are independently hydrogen, —OH, substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl, or substituted orunsubstituted heteroaryl; where R¹ and R² may optionally be joined toform a substituted or unsubstituted heterocycloalkyl or a substituted orunsubstituted heteroaryl.

2. The compound, or a pharmaceutically acceptable salt thereof, ofembodiment 1, wherein the compound, or a pharmaceutically acceptablesalt thereof, is not

3. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 1 to 2, wherein the compound has a formula selectedfrom the group consisting of:

4. The compound of any one of embodiments 1 to 3, or a pharmaceuticallyacceptable salt thereof, wherein L⁰ is a bond.

5. The compound of any one of embodiments 1 to 3, or a pharmaceuticallyacceptable salt thereof, wherein L⁰ is an unsubstituted C₁-C₆ alkylene.

6. The compound of any one of embodiments 1 to 3, or a pharmaceuticallyacceptable salt thereof, wherein L⁰ is an unsubstituted C₁-C₄ alkylene.

7. The compound of any one of embodiments 1 to 3, or a pharmaceuticallyacceptable salt thereof, wherein L⁰ is an unsubstituted methylene.

8. The compound of any one of embodiments 1 to 7, or a pharmaceuticallyacceptable salt thereof, wherein R¹ is L¹-R³; L¹ is independently a bondor an unsubstituted C₁-C₁₀ alkylene; and R³ is independently hydrogen,

halogen, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂,—NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂,—NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H,—NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, or substituted or unsubstitutedheteroaryl.

9. The compound of any one of embodiments 1 to 8, or a pharmaceuticallyacceptable salt thereof, wherein L¹ is a bond.

10. The compound of any one of embodiments 1 to 8, or a pharmaceuticallyacceptable salt thereof, wherein L¹ is an unsubstituted C₁-C₄ alkylene.

11. The compound of any one of embodiments 1 to 8, or a pharmaceuticallyacceptable salt thereof, wherein L¹ is an unsubstituted methylene.

12. The compound of any one of embodiments 1 to 11, or apharmaceutically acceptable salt thereof, wherein R³ is independentlyhydrogen, halogen, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH,—CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H,—SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H,—NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂, 4-methoxybenzyl,4-methoxyphenyl, or pyridyl.

13. The compound of any one of embodiments 1 to 12, or apharmaceutically acceptable salt thereof, wherein R² is L²-R⁴; L² isindependently a bond or an unsubstituted C₁-C₁₀ alkylene; and R⁴ isindependently hydrogen,

halogen, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂,—NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂,—NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H,—NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, or substituted or unsubstitutedheteroaryl.

14. The compound of any one of embodiments 1 to 13, or apharmaceutically acceptable salt thereof, wherein L² is a bond.

15. The compound of any one of embodiments 1 to 13, or apharmaceutically acceptable salt thereof, wherein L² is an unsubstitutedC₁-C₄ alkylene.

16. The compound of any one of embodiments 1 to 13, or apharmaceutically acceptable salt thereof, wherein L² is an unsubstitutedmethylene.

17. The compound of any one of embodiments 1 to 16, or apharmaceutically acceptable salt thereof, wherein R⁴ is independentlyhydrogen, halogen, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH,—CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H,—SO₂NH₂, —NHNH₂, —OCH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H,—NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂, 4-methoxybenzyl,4-methoxyphenyl, or pyridyl.

18. The compound of any one of embodiments 1 to 7, or a pharmaceuticallyacceptable salt thereof, wherein R¹ and R² are joined to form anunsubstituted heterocycloalkyl.

19. The compound of any one of embodiments 1 to 7, or a pharmaceuticallyacceptable salt thereof, wherein R¹ and R² are joined to form anunsubstituted piperidinyl.

20. The pharmaceutically acceptable salt of any one of embodiments 1 to19, wherein the compound has a formula selected from the groupconsisting of formula I, II, IV, Ia, IIa, and IVa, and wherein thecompound comprises a protonated nitrogen cation.

21. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 1 to 2, wherein the compound is selected from thegroup consisting of

22. A pharmaceutical composition comprising a pharmaceuticallyacceptable excipient and a compound, or pharmaceutically acceptable saltthereof, of any one of embodiments 1 to 21.

23. The pharmaceutical composition of embodiment 22, comprising a secondagent wherein said second agent is an anti-cancer agent.

24. A method of treating cancer in a patient in need of said treatment,said method comprising administering a therapeutically effective amountof a compound of any one of embodiments 1 to 21, or a pharmaceuticallyacceptable salt thereof, to said patient.

25. The method of embodiment 24, wherein said cancer is breast cancer.

26. The method of embodiment 25, wherein said breast cancer is resistantto an anti-cancer agent.

27. The method of embodiment 26, wherein said anti-cancer agent istrastuzumab.

28. The method of embodiment 25, wherein said breast cancer is resistantto a hormonal therapeutic agent.

29. The method of embodiment 28, wherein said hormonal therapeutic agentis tamoxifen.

30. The method of embodiment 24, wherein said cancer is lung cancer.

31. The method of embodiment 30, wherein said lung cancer is non-smallcell lung cancer.

32. The method of any one of embodiments 30 to 31, wherein said lungcancer is resistant to an anti-cancer agent.

33. The method of embodiment 32, wherein said anti-cancer agent is aplatinum-based compound.

34. The method of any one of embodiments 32 to 33, wherein saidanti-cancer agent is cisplatin or carboplatin.

35. A method of treating a non-malignant hyperproliferative disease in apatient in need of said treatment, said method comprising administeringa therapeutically effective amount of a compound of any one ofembodiments 1 to 21, or a pharmaceutically acceptable salt thereof, tosaid patient.

36. The method of embodiment 35, wherein said non-malignanthyperproliferative disease is selected from the group consisting of ahamartomatous lesion, angiomyolipoma, lymphangioleiomyomatosis, tuberoussclerosis complex, a hamartia, or a hamartoma.

37. A method of inhibiting cancer cell growth or survival comprisingcontacting the cell with an effective amount of a compound of any one ofembodiments 1 to 21, or a pharmaceutically acceptable salt thereof.

38. The method of embodiment 37, wherein said cancer cell is a cancerstem cell.

39. The method of embodiment 37, wherein said cancer cell is a cancerprogenitor cell.

40. The method of any one of embodiments 37 to 38, wherein said cancercell is a lung cancer cell or breast cancer cell.

41. The method of embodiment 37, wherein said compound, or apharmaceutically acceptable salt thereof, increases apoptosis of saidcancer cells.

42. A method of decreasing the level of activity of NF-κB in a cellcomprising contacting the cell with an effective amount of a compound ofany one of embodiments 1 to 21, or a pharmaceutically acceptable saltthereof.

43. A method of increasing the level of activity of TSC1 in a cellcomprising contacting the cell with an effective amount of a compound ofany one of embodiments 1 to 21, or a pharmaceutically acceptable saltthereof.

44. A method of increasing the level of activity of TSC2 in a cellcomprising contacting the cell with an effective amount of a compound ofany one of embodiments 1 to 21, or a pharmaceutically acceptable saltthereof.

45. A method of decreasing the level of activity of mTOR in a cellcomprising contacting the cell with an effective amount of a compound ofany one of embodiments 1 to 21, or a pharmaceutically acceptable saltthereof.

VII. Examples

The following examples are offered to illustrate, but not to limit theclaimed invention.

A. ISOLATION OF CANCER STEM/PROGENITOR CELLS FROM LUNG CANCER CELL LINES

Substantial evidence has accumulated to support the role of a smallsubset of self-renewing cells that sustain malignant growth. Thissubpopulation is termed cancer-initiating cells or cancerstem/progenitor cells (CSC) for their high capacity for self-renewal andsuperior levels of malignancy. Cancer stem cells are identified andisolated in several malignancies including lung cancer. CD133(prominin-1), a 5-transmembrane glycoprotein, has been reported as animportant biomarker to identify subsets of human lung CSC. Further,isolated lung cancer cells with high aldehyde dehydrogenase-1 (ALDH)activity display in vitro and in vivo features of cancer stem cells andexpress surface marker CD133. Based on these findings, we identifiedCD133+/ALDH+ tumor stem/progenitor cells from human lung cancer cells invitro using an Aldefluor assay (to isolate ALDH+ cells) (5,7,8) incombination with labeled anti-CD133 antibodies (3,10) followed byfluorescence-activated cell sorting analysis (11). In brief, ALDHactivity and CD133 expression were analyzed by established doublelabeling methods using the Aldefluor Assay Kit (Stem Cell Technologies)and CD133/1 antibody (Miltenyi) according to the manufacturer'sinstructions. Cells were suspended in assay buffer and incubated withthe ALDH substrate Aldefluor or with Aldefluor and the ALDH inhibitordiethylaminobenzaldehyde DEAB (negative control) for 30 min at 37 C.Cells were subsequently washed and labeled with Allophycocyanin(APC)-conjugated CD133/1 for 30 min on ice. Cells were washed with assaybuffer twice and analyzed using FACSCalibur (BD Bioscience) andCellQuest software (BD Bioscience). Data were analyzed using FlowJo(Tree Star Inc.). Treatment with estrogen, a known risk factor for lungcancer progression (12), stimulated a modest increase in the numbers ofCSC (FIG. 1). This latter effect of estrogens was blocked by use ofantiestrogens (11).

B. PROPERTIES OF ISOLATED CANCER STEM CELLS

We cultivated subpopulations of lung tumor cells isolated followingtreatment with steroid hormones (11) and determined that theseCD133+/ALDH+ subsets were able to grow as tumor spheres and maintainself-renewal capacity, properties indicative of CSC. In contrast,control CD133−/ALDH− subsets were not capable of significant tumorsphere formation (FIG. 2).

Further in vitro studies of CSC subsets indicate that these cells aremore resistant to cisplatin and docetaxel chemotherapy treatment thanbulk NSCLC cells. Moreover, the CSC subpopulations exhibit constituitivenuclear localization of NFκB as determined by immunofluorescencemicroscopy.

C. PARTHENOLIDE ANALOGUES INHIBIT GROWTH OF LUNG CANCER STEM CELLS

Analogues of parthenolide were prepared and purified (FIG. 8) andassessed for antitumor efficacy, to develop the best agents for clinicaluse. Responses of CSC-like subsets, bulk tumor cells and normal lungepithelial cells to parthenolide (PTL) analogues alone and withchemotherapy are ongoing, with assessments by TUNEL and Annexin-V assayfor apoptosis (13) (FIG. 3) and cell proliferation (FIG. 4).

To assess drug interactions in vitro and statistical significance,t-tests, ANOVA or other appropriate tests were used (5,14,15). Anexample of results with parthenolide analogue JD201 (DMAPT) is shown inFIG. 3 (apoptosis data) and FIG. 4 (cell proliferation).

These findings using JD201 (DMAPT) are promising for antitumor activityin NSCLC cells, particularly in CSC subsets.

D. EFFECT OF PARTHENOLIDE ANALOGUES IN BULK NON-SMALL CELL LUNG CANCERCELL LINES

To determine the effect of parthenolide analogues in lung cancer cellproliferation we tested two male (A549, H23) and two female (H2122,H1975) lung cancer cell lines with increasing concentrations ofparthenolide analogues (0.1, 1, 10 and 100 μM) for 72 hours. FIG. 5shows growth inhibition by compounds 201-210 in a dose dependent manner(P>0.01). Significant growth inhibition was observed at 10 μM (201, 202and 203) and 100 μM (201, 202, 203, 205 and 207) concentrations ofparthenolides. Data shown in FIG. 5 indicate that parthenolide analoguesJD 201, 202 and 203 had greater inhibitory activity than compounds 205and 207.

E. PARTHENOLIDE ANALOGUES SENSITIZE LUNG TUMOR CELLS TOCISPLATIN-INDUCED CYTOTOXICITY

In addition, to determine if parthenolides can sensitize lung cancercells to cytotoxicity induced by cisplatin, a drug commonly used in theclinic, we treated cells with 10 and 50 μM concentrations ofparthenolide analogues in the presence of 5 μM cisplatin, a suboptimaldose that does not elicit significant cytotoxicity when administeredalone. After 72 hours, LDH release was measured as a correlate of celldeath and membrane disruption. Exposure to 10 μM cisplatin is shown asan independent control of cytotoxicity (reduced cell viability) inducedby cisplatin. Compound JD201 significantly increased the cytotoxicityinduced by cisplatin in both NSCLC cell lines assessed in these studies(see FIG. 6).

Upon confirming that parthenolide analogues can inhibit NSCLC cellproliferation, induce cell death and enhance the cytotoxicity ofchemotherapy, we began to investigate potential cellular mechanisms thatmay underlie this process. For example, we used Western Blot methods todetermine phosphorylation of the p65 subunit of NF-κB. H23 cells wereincubated in the presence of vehicle control or 10 μM concentration ofthe parthenolide derivatives (JD 201, 202, 203, 204) for 60 min. Aftertreatment, cells were collected and homogenized in lysis buffer.Extracts were electrophoresed on 4-20% gradient SDS-PAGE gels andtransferred to nitrocellulose membranes. Blots were probed withanti-phospho-NF-κB/p65 and NF-κB/p65 for a loading control. Incubationof cells in the presence of parthenolides reduced phosphorylation of thep65 subunit of NF-κB (See FIG. 7).

To further confirm the tumorigenic potential of CSC subsets(CD133+/ALDH+ cells) as compared with non-CSC populations (CD133−/ALDH−cells), we implanted the tumor cell subpopulations subcutaneously inimmunosuppressed mice [16]. Tumor growth occurred in all 3 of 3 miceinoculated subcutaneously with either 200 or 20,000 CSC (CD133+/ALDH+cells). In contrast, no tumor growth was detected with inoculation of200 non-CSC (CD133−/ALDH− tumor cells), but 1 of 3 mice developed tumorswith injection of 20,000 non-CSC (CD133−/ALDH− cells) (16). Thus, thesefindings suggest that CSC (CD133+/ALDH+ tumor cells) show a greatertumorigenic capability than non-CSC subsets (CD133−/ALDH− cell) in vivo.Studies of the effects of a parthenolide analogue alone and incombination with chemotherapy are in progress. Initial in vivo findingswith A549 NSCLC xenografts implanted in immunodeficient mice indicatethat parthenolide analogue JD201 elicits a reduction in tumorprogression when administered as an oral gavage (100 mg/kg/day) for 21days as compared to a vehicle control, with tumor volumes of 55±5 mm³ inthe JD201 group versus 252±31 mm³ in controls. Additional studies of theinteraction of this drug with chemotherapy in vivo are continuing.

We have identified subpopulations of chemotherapy-resistant human NSCLCcells with enrichment for CSC biomarkers and significant CSC activity.To target cancer stem cells and suppress tumor progression, wesynthesized and tested analogues of parthenolide that show stronganticancer activity, with marked suppression of CSC subsets. Use ofparthenolide analogues may lead to a new strategy to eradicate lungcancer.

F. GENERAL METHODS FOR BREAST CANCER EXPERIMENTS

Studies detailed below were done primarily using a panel of MCF-7 andSKBR3 human breast cancer cells (ATCC) with different properties.Overall, this panel include:

TABLE 1 1. Cell Line Cell Properties MCF-7/CON MCF-7 parent breast tumorcells without HER2 overexpression and ER+ (2) MCF-7/HER2 MCF-7 breasttumor cells with HER2 overexpression and ER+ (2) MCF-7/TMR MCF-7 breasttumor cells with acquired tamoxifen resistance and ER+ (38) SKBR3/CONSKBR3 parent breast tumor cells without HER2 overexpression and ER−SKBR3/TZR SKBR3 breast tumor cells with HER2 overexpression and acquiredtraztuzumab (Herceptin) resistance (39)

MCF-7/CON cells are tamoxifen-sensitive, while MCF-7/HER2 and MCF-7/TMRcells are known to be tamoxifen-resistant (28,61,66,67). Except for TMR(64,68), cells were routinely cultured in RPMI 1640 media with 10%heat-inactivated FBS (28,64,66). For E2-free conditions, media werechanged 48 h before studies to phenol-red free RPMI 1640 with 1%dextran-coated, charcoal-treated (DCC) FBS. TMR cells were derived bylong-term continuous exposure of cells to 10⁻⁷ M 4-hydroxy-tamoxifen byestablished methods (64,68) and were maintained with 10⁻⁷ M4-hydroxytamoxifen in medium. Levels of ERα and HER2 expression in theseseveral cell lines were determined by use of Western blot methods. SKBR3cells resistant to trastuzumab were developed and maintained as detailedbefore (65)

Cell Transfection and NF-kappaB Activity. Activity of NFkB was assessedby using established transient transfection and luciferase reporterassays. Cell cultures were seeded one day before transfection withluciferase (luc) reporters to a density of 1-2×10³ cells/well in 96-wellplates and using established growth media (67,69). Cells weretransiently transfected with 0.5 μg of NFκB-luc reporter vector(Promega), along with FuGene 6 transfection reagent (Roche). Renillaluciferase vector pRL-tk-luc (Promega) was co-transfected as an internalcontrol reporter vector to normalize for transfection efficiency.Culture media was changed 20 h following transfection and cells werethen maintained in appropriate media for 24 h. As per manufacturer'srecommendations, cells were subsequently washed with PBS, lysed forDual-Glo luciferase assay (Promega), and reporter activity measured byluminometer. The ratio of firefly luminescence/Renilla luminescence wasused for comparison of NFκB driven gene activities in control cells.

Assays for Tumor Cell Proliferation.

Breast tumor cells were be grown in RPMI 1640 media with 10%heat-inactivated FBS (except for TMR and TZR cells as noted above). Forestrogen-free conditions, medium were changed 48 h before studies tophenol-red free RPMI 1640 with 1% dextran-coated, charcoal-treated (DCC)FBS, then treated with selected reagents. Cell counts and viabilitytests (Trypan blue) will be done every 24 hr for 2-33 days. After 48-72hr, proliferation was assessed by cell counts.

Statistical Analyses.

For this in vitro work, triplicates of experiments were done. Student'st-test, ANOVA, or Kruskal-Wallis test if outcomes are non-normallydistributed, were used as appropriate to compare intervention groups.Analyses were evaluated using bar and scatter graphs with means, SD, SE.Time trend curves for agents under different conditions were obtained asappropriate. Repeated measures ANOVA was used as needed to assess time,condition, time by condition interaction effects. P<0.05 was consideredsignificant.

G. ENDOCRINE RESISTANT BREAST CANCER

NFkB activity is enhanced in endocrine-resistant breast tumors. Toinvestigate NFκB activation in endocrine-resistant breast cancer cells,we used tumor cell lines with known resistance to tamoxifen. For thesestudies, we used MCF-7 human breast cancer cells that express abundantERα and are sensitive to tamoxifen (28,29,67). The activity of NFkB inMCF-7 control cells was compared to that of tamoxifen-resistant MCF-7(MCF-7/TMR) and MCF-7 HER2-overexpressing (MCF-7/HER2) cells (see FIG.12).

Parthenolide analogues reduce proliferation of breast cancer cells withtamoxifen resistance. To investigate the effects of parthenolide (PTL)analogues on breast tumor cell proliferation in vitro, we cultivatedMCF-7 breast cancer cells with HER-2 overexpression (28) in the presenceand absence of PTL analogues 201 and 203 (see FIG. 13). As noted above,these MCF-7/HER2 cells are tamoxifen-resistant. Both analogues eliciteda dose-dependent reduction in proliferation of MCF-7/HER2 cells ascompared to controls.

Parthenolide analogue enhances antitumor efficacy of tamoxifen intamoxifen-resistant MCF-7/HER2 breast cancer cells. To further assessthe activity of parthenolide analogue 203 when administered incombination with tamoxifen, we used MCF-7/HER2 cells with establishedtamoxifen-resistance (see FIG. 14). The results of this study indicatethat parthenolide analogue 203 may help to sensitize theseendocrine-resistant cells to tamoxifen.

Parthenolide analogue promotes antitumor effects of tamoxifen intamoxifen-resistant MCF 7/TAMR breast cancer cells. To further assessthe activity of parthenolide analogues in reversing tamoxifen resistancein human breast cancer cells, we used tamoxifen-resistant (TAM-R) cellsdeveloped as described previously (64,68). (see FIG. 14).

Results of these studies show that the MCF-7/TMR cells are resistant totamoxifen treatment in the absence of parthenolide analogues. However,combination of tamoxifen with parthenolide analogue 203 elicits enhancedblockade of tumor cell proliferation over a wide dose range of theanalogue as compared to appropriate controls (P<0.01).

H. HER2+ BREAST CANCERS WITH RESISTANCE TO TRASTUZUMAB

Activation of NFkB in HER2-overexpressing breast tumor cells resistantto Herceptin (trastuzumab) and effects of parthenolide analogues. Thehuman breast cancer cell line SKBR3 (ATCC) was obtained from theAmerican Type Culture Collection (ATCC). Unless otherwise stated,monolayer cultures of SKBR3 were maintained in Dulbeccos' ModifiedEagle's Media (DMEM):F12 medium with 10% FBS. The cell linesoverexpressed the HER2 gene product. Trastuzumab-resistant clonesSKBR3/TZR were generated from SKBR3 cells as detailed previously (65).In brief, to select trastuzumab-resistant clones, SKBR3 cells wereplated in 24-well plates at low density and maintained in growth mediumcontaining 10, 50, and 100 mg/mL of trastuzumab. SKBR3/TZR cells weremaintained in growth medium containing 100 mg/mL of trastuzumab.SKBR3/TZR cells were repeatedly confirmed as insensitive to trastuzumab.Data from routine proliferation assays showed that growth of SKBR3parental (control) cells was significantly inhibited by fresh treatmentwith 10 and 100 mg/mL of trastuzumab, whereas the growth of SKBR3/TZRcells which were maintained in media containing 100 mg/mL trastuzumabwere not inhibited in 2-5 days. The number of cells overexpressing HER2receptors was reduced in SKBR3 cells on treatment with trastuzumab (10mg/mL) for 72 hrs. However, SKBR3/TZR cells maintained in growth mediacontaining 100 mg/mL trastuzumab for extended periods of time did notexhibit any significant difference in overall HER2 receptoroverexpression compared with untreated parental cells.

Results of experiments to assess NFkB activity in SKBR3 cells thatsensitive (SKBR3/CON) and resistant to traztuzumab (SKBR3/TZR) treatmentin vitro are shown in FIG. 8. These experiments indicate that oneconsequence of trastuzumab resistance in HER2+ breast cancer cells isthe enhancement of NFkB activity. The data in FIG. 17 show thatSKBR3/TZR cells have significantly increased NFkB activity as comparedto SKBR3/CON cells (P<0.001). Such findings correlate with independentobservations on the essential role of NFκB in HER2-induced oncogenesisby providing signals that maintain mammary tumor-initiating cells. Thenotion that HER2 can regulate NFκB and that NFκB can in turn regulatedownstream pathways that, in turn, regulate HER2 suggests that these twoimportant stem cell regulatory genes may interact in controlling cancerprogression (70).

To determine the antitumor efficacy of parthenolide analogues known toblock NFkB activation, we assessed their effects on proliferation ofSKBR3/TZR cells in vitro (FIG. 17). These results show that parthenolideanalogues elicit significant growth inhibition in HER2-overexpressingcells that are trastuzumab-resistant either when administered alone orin combination with trastuzumab (P<0.001). Further investigation of thisunique strategy to block the progression of trastuzumab(Herceptin)-resistant breast cancers has important implications for themanagement of patients afflicted with this disease in the clinic.

I. PARTHENOLIDES IN NONMALIGNANT PROLIFERATIVE DISEASE

We have tested inhibitory activity of parthenolide analogues using cellmodels for TSC-related proliferative disease. The ELT3 cells are aTSC2-null smooth muscle-derived cell line from a rat uterine leiomyomaand often used as a model for lymphangioleiomyomatosis (LAM) and otherTSC-related proliferative diseases (75,76). Angiomyolipoma cells (AML)that have inactivating mutations in both alleles of the TSC2 gene werealso used as an alternate cell model (77,78). Cells were routinelycultured in IIA complete medium (50% DMEM, 50% F-12, 1.2 g/ml NaHCO3,1.6 mM FeSO4, 50 nM sodium selenite, 25 mg/ml insulin, 200 nMhydrocortisone, 10 mg/ml transferrin, 1 nM triiodothyronine, 10 mU/mlvasopressin, 10 nM cholesterol, 10 ng/ml epidermal growth factor)containing 15% FBS. For study of proliferation, cells were incubated for3 days in IIA medium containing 1% FBS. Effects of parthenolide analogueJD203 on proliferation of ELT3 and AML cells in vitro are shown in FIG.19. Results of these experiments indicate that parthenolides may alsohave utility in the treatment of TSC-related proliferative diseases thatare not classified as malignancies.

J. GENERAL EXPERIMENTAL DETAIL FOR CHEMICAL SYNTHESIS OF PARTHENOLIDEANALOGUES

Dichloromethane and methanol were distilled from calcium hydride underan argon atmosphere. Diethyl ether was distilled from benzoquinone ketylradical under an argon atmosphere. All other solvents or reagents werepurified according to literature procedures.

High resolution mass spectrometry data was obtained on a Waters LCTPremier XE Time of Flight LC-MS. ¹H and ¹³C NMR spectra were obtained onAV-300, ARX-400, ARX-500 or Avance-500 spectrometers. The chemicalshifts are reported in parts per million (ppm, 6). The couplingconstants are reported in Hertz (Hz) and the resonance patterns arereported with the following notations: br (broad), s (singlet), d(doublet), t (triplet), q (quartet) and m (multiplet). Thin-layerchromatography (TLC) was carried out using precoated silica gel sheets(Merck 60 F₂₅₄). Visual detection was performed with ultraviolet light(short wave and long wave), p-anisaldehyde stain, and potassiumpermanganate stain. Flash chromatography was performed usingSilicaFlash™ P60 (60 A, 40-63 mm) silica gel from SiliCycle Inc. withcompressed air.

General Procedure A:

For Compounds Dg-2-11, Dg-2-12, Dg-2-13, Dg-2-36b, Dg-2-75. Parthenolide(20 mg, 0.08 mmol) and an excess of the amine (0.24 mmol) were dissolvedin methanol (3.0 mL) and the mixture was stirred under argon at 21° C.overnight. The reaction mixture was extracted with ethyl acetate (3×15mL), washed with water and brine, and dried over MgSO₄. After removal ofthe solvent by rotary evaporation, the crude product was purified byflash column chromatography to obtain the desired dialkylaminopathenolide adducts.

General Procedure B:

For Compounds JD201, JD202, JD203, JD204, JD207. The dialkylaminoparthenolide adduct from above (0.01 mmol) was added to the stirredsolution of fumaric acid (0.005 M, 2.0 mL) in diethyl ether at 0° C.After the mixture stirred for 10 min at 0° C., the reaction temperaturewas raised to 21° C. and the mixture was stirred for 2 h. The solventwas removed by rotary evaporation to afford the desired compounds.

(1aR,7aS,8R,10aS,Z)-8-((Dimethylamino)methyl)-1a,5-dimethyl-2,3,6,7,7a,8,10a,10b-octa-hydrooxireno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one

The title compound, Dg-2-11, was obtained by following general procedureA using dimethylamine in 98% yield after flash column chromatography(eluent: dichloromethane/MeOH=20/1). ¹H NMR (300 MHz, CDCl₃): δ 5.19(1H, d, J=9.6 Hz), 3.82 (1H, t, J=9.0 Hz), 2.27 (6H, s), 1.68 (3H, s),1.28 (3H, s). ¹³C NMR (75 MHz, CDCl₃): δ 176.4, 134.6, 125.0, 82.1,66.4, 61.4, 57.5, 47.9, 46.4, 46.0, 41.0, 36.6, 29.9, 24.1, 17.2, 16.9.HR-MS (ESI) Calcd for [C₁₇H₂₇NO₃+H]⁺ 294.2069. found 294.2073.

(1aR,7aS,8R,10aS,Z)-8-((Dimethylamino)methyl)-1a,5-dimethyl-2,3,6,7,7a,8,10a,10b-octa-hydrooxireno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one,fumaric acid salt, JD201

The title compound, JD201, was obtained by following general procedureB. ¹H NMR (400 MHz, DMSO-d₆): δ 6.61 (2H, s), 5.22 (1H, d, J=12.0 Hz),3.98 (1H, t, J=12.0 Hz), 2.24 (6H, s), 1.65 (3H, s), 1.21 (3H, s). ¹³CNMR (75 MHz, CDCl₃): δ 177.1, 166.6, 134.8, 134.5, 125.0, 81.9, 65.8,61.6, 57.7, 47.7, 46.0, 45.5, 40.9, 36.5, 29.1, 24.1, 17.3, 17.0. HR-MS(ESI) Calcd for [C₁₇H₂₇NO₃+H]⁺ 294.2069. found 294.2058.

(1aR,7aS,8R,10aS,Z)-1a,5-Dimethyl-8-(((pyridin-2-ylmethyl)amino)methyl)-2,3,6,7,7a,8,10a,10b-octahydrooxireno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one

The title compound, Dg-2-13, was obtained by following general procedureA using 2-(aminomethyl)pyridine in 97% yield after flash columnchromatography (eluent: dichloromethane/MeOH=20/1). ¹H NMR (300 MHz,CDCl₃): δ 8.52 (1H, d, J=4.8 Hz), 7.64 (1H, dt, J=1.8, 7.8 Hz), 7.33(1H, d, J=7.8 Hz), 7.16 (1H, d, J=5.4 Hz), 5.14 (1H, d, J=11.4 Hz), 4.09(1H, dd, J=6.0, 15.0 Hz), 3.94 (1H, dd, J=15.0, 21.0 Hz), 3.83 (1H, t,J=9.0 Hz), 1.66 (3H, s), 1.26 (3H, s). ¹³C NMR (75 MHz, CDCl₃): δ 176.6,159.3, 149.1, 136.5, 134.4, 125.0, 122.1, 122.0, 82.5, 66.2, 61.4, 55.1,47.9, 46.6, 46.5, 40.9, 36.5, 29.8, 24.0, 17.1, 16.8. HR-MS (ESI) Calcdfor [C₂₁H₂₈N₂O₃+H]⁺ 357.2178. found 357.2191.

(1aR,7aS,8R,10aS,Z)-1a,5-Dimethyl-8-(((pyridin-2-ylmethyl)amino)methyl)-2,3,6,7,7a,8,10a,10b-octahydrooxireno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one,fumaric acid salt, JD202

The title compound, JD202, was obtained by following general procedureB. ¹H NMR (400 MHz, DMSO-d₆): δ 8.49 (1H, d, J=4.0 Hz), 7.74 (1H, t,J=7.5 Hz), 7.41 (1H, d, J=7.9 Hz), 7.24 (1H, dd, J=4.8, 7.4 Hz), 6.57(2H, s), 5.17 (1H, d, J=12.5 Hz), 3.97 (1H, t, J=9.4 Hz), 1.60 (3H, s),1.16 (3H, s). ¹³C NMR (75 MHz, CDCl₃): δ 176.6, 166.1, 158.9, 148.8,136.6, 134.4, 134.1, 124.5, 122.2, 122.0, 81.7, 65.4, 61.2, 54.2, 47.1,45.7, 45.6, 40.5, 36.1, 28.9, 23.7, 16.8, 16.6. HR-MS (ESI) Calcd for[C₂₁H₂₈N₂O₃+H]⁺ 357.2178. found 357.2191.

(1aR,7aS,8R,10aS,Z)-8-((Bis(2-hydroxyethyl)amino)methyl)-1a,5-dimethyl-2,3,6,7,7a,8,10a,10b-octahydrooxireno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one,Dg-2-12

The title compound, Dg-2-12, was obtained by following general procedureA using diethanolamine in 98% yield after flash column chromatography(dichloromethane/MeOH=20/1). ¹H NMR (300 MHz, CDCl₃): δ 5.17 (1H, d,J=11.7 Hz), 3.87 (1H, t, J=9.0 Hz), 1.67 (3H, s), 1.27 (3H, s). ¹³C NMR(75 MHz, CDCl₃): δ 177.8, 134.3, 125.1, 82.4, 66.2, 61.6, 59.2, 59.2,56.5, 56.5, 53.0, 47.3, 46.3, 40.9, 36.5, 29.7, 24.0, 17.1, 16.8. HR-MS(ESI) Calcd for [C₁₉H₃₁NO₅+H]⁺ 354.2281. found 354.2294.

(1aR,7aS,8R,10aS,Z)-8-((Bis(2-hydroxyethyl)amino)methyl)-1a,5-dimethyl-2,3,6,7,7a,8,10a,10b-octahydrooxireno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one,fumaric acid salt, JD203

The title compound, JD203, was obtained by following general procedureB. ¹H NMR (500 MHz, DMSO-d₆): δ 6.61 (2H, s), 5.18 (1H, d, J=12.4 Hz),3.94 (1H, t, J=9.5 Hz), 1.63 (3H, s), 1.18 (3H, s). ¹³C NMR (125 MHz,DMSO-d₆): δ 177.4, 166.3, 134.9, 134.3, 124.5, 82.0, 65.9, 61.5, 59.4,59.4, 57.2, 57.2, 54.0, 46.7, 46.4, 40.0, 36.5, 29.2, 24.0, 17.2, 17.0.HR-MS (ESI) Calcd for [C₁₉H₃₁NO₅+H]⁺ 354.2281. found 354.2284.

(1aR,7aS,8R,10aS,Z)-8-((Methoxy(methyl)amino)methyl)-1a,5-dimethyl-2,3,6,7,7a,8,10a,10b-octahydrooxireno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one,Dg-2-36b

The title compound, Dg-2-36b, was obtained by following generalprocedure A using N,O-dimethyl-hydroxylamine hydrochloride and potassiumcarbonate (33.2 mg, 0.24 mmol) in ethanol as the solvent in 79% yieldafter flash column chromatography (eluent: hexane/ethyl acetate=4/1).The ethoxy adduct, JD206, was also obtained in 8% yield in thisreaction. ¹H NMR (300 MHz, CDCl₃): δ 5.19 (1H, d, J=11.1 Hz), 3.83 (1H,t, J=9.0 Hz), 3.50 (3H, s), 2.63 (3H, s) 1.71 (3H, s), 1.30 (3H, s). ¹³CNMR (75 MHz, CDCl₃): δ 176.0, 134.7, 125.0, 82.0, 66.6, 61.4, 59.0,58.4, 47.5, 46.2, 45.0, 41.1, 36.7, 29.8, 24.1, 17.2, 17.0. HR-MS (ESI)Calcd for [C₁₇H₂₇NO₄+H]⁺ 310.2018. found 310.2026.

(1aR,7aS,8R,10aS,Z)-8-((Methoxy(methyl)amino)methyl)-1a,5-dimethyl-2,3,6,7,7a,8,10a,10b-octahydrooxireno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one,fumaric acid salt, JD204

The title compound, JD204, was obtained by following general procedureB. ¹H NMR (300 MHz, MeOD): δ 6.75 (2H, s), 5.25 (1H, d, J=9.6 Hz), 4.00(1H, t, J=9.0 Hz), 3.49 (3H, s), 2.60 (3H, s) 1.72 (3H, s), 1.30 (3H,s). ¹³C NMR (75 MHz, MeOD): δ 179.1, 168.0, 168.0, 136.2, 135.2, 135.2,125.8, 83.6, 68.1, 63.2, 59.4, 59.3, 48.2, 47.2, 45.2, 42.0, 37.7, 30.5,25.0, 17.5, 17.1. HR-MS (ESI) Calcd for [C₁₇H₂₇NO₄+H]⁺ 310.2018. found310.2031.

(1aR,7aS,8R,10aS,Z)-8-((Hydroxy(methyl)amino)methyl)-1a,5-dimethyl-2,3,6,7,7a,8,10a,10b-octahydrooxireno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one,Dg-2-75

The title compound, Dg-2-75, was obtained by following general procedureA using N-methylhydroxyl-amine hydrochloride and potassium carbonate(33.2 mg, 0.24 mmol) in 93% yield after flash chromatography (eluent:dichloromethane/MeOH=40/1). ¹H NMR (300 MHz, CDCl₃): δ 5.17 (1H, d,J=11.7 Hz), 3.84 (1H, t, J=9.0 Hz), 2.70 (3H, s) 1.68 (3H, s), 1.28 (3H,s). ¹³C NMR (75 MHz, CDCl₃): δ 176.8, 134.6, 124.9, 82.3, 66.3, 61.6,59.7, 49.5, 47.5, 45.9, 41.0, 36.6, 29.6, 24.0, 17.1, 16.9. HR-MS (ESI)Calcd for [C₁₆H₂₅NO₄+H]⁺ 296.1862. found 296.1859.

(1aR,7aS,8R,10aS,Z)-8-((Hydroxy(methyl)amino)methyl)-1a,5-dimethyl-2,3,6,7,7a,8,10a,10b-octahydrooxireno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one,fumaric acid salt, JD207

The title compound, JD207, The title compound was obtained by followinggeneral procedure B. ¹H NMR (300 MHz, MeOD): δ 6.75 (2H, s), 5.25 (1H,d, J=9.9 Hz), 3.98 (1H, t, J=9.0 Hz), 2.68 (3H, s) 1.72 (3H, s), 1.30(3H, s). ¹³C NMR (75 MHz, MeOD): δ 179.3, 168.2, 168.2, 136.3, 135.2,135.2, 125.7, 83.9, 68.0, 63.2, 60.1, 49.7, 47.9, 47.1, 41.9, 37.7,30.5, 25.0, 17.5, 17.1. HR-MS (ESI) Calcd for [C₁₆H₂₅NO₄+H]⁺ 296.1862.found 296.1864.

(1aR,7aS,8R,10aS,Z)-8-(Ethoxymethyl)-1a,5-dimethyl-2,3,6,7,7a,8,10a,10b-octahydrooxi-reno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one,JD206

The title compound, JD206, was obtained in the reaction that producedcompound Dg-2-36b above in 8% yield. ¹H NMR (300 MHz, CDCl₃): δ 5.22(1H, d, J=9.9 Hz), 3.83 (1H, t, J=9.0 Hz), 3.75 (1H, dq, J=3.0, 9.0 Hz),3.54 (1H, dq, J=3.0, 9.0 Hz), 2.77 (1H, d, J=9.0 Hz), 2.41 (1H, d, J=9.0Hz), 1.70 (3H, s), 1.25 (3H, s), 1.19 (3H, t, J=9.0 Hz). ¹³C NMR (75MHz, CDCl₃): δ 175.1, 134.6, 125.1, 82.2, 67.0, 66.4, 66.3, 61.4, 48.6,45.8, 41.1, 36.6, 30.0, 24.1, 17.2, 16.9, 15.0. HR-MS (ESI) Calcd for[C₁₇H₂₆O₄+H]⁺ 295.1909. found 295.1918.

(1aR,7aS,8R,10aS,Z)-8-(((4-methoxybenzyl)oxy)methyl)-1a,5-dimethyl-2,3,6,7,7a,8,10a,10b-octahydrooxireno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one,JD208

Parthenolide (49 mg, 0.2 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene(DBU, 0.3 μL, 2.0 mmol) were dissolved in 4-methoxybenzyl alcohol (1.0mL), and the mixture was stirred under argon at 21° C. overnight. Thereaction mixture was extracted with ethyl acetate (3×15 mL), washed withwater and brine, and dried over MgSO₄. After removal of the solvent byrotary evaporation, the crude product was purified by flash columnchromatography (eluent: hexane/ethyl acetate=4/1) to obtain the desiredproduct, JD208. ¹H NMR (300 MHz, CDCl₃): δ 7.24 (2H, d, J=8.4 Hz), 6.88(2H, d, J=8.4 Hz), 5.17 (1H, d, J=11.4 Hz), 4.49 (2H, q, J=11.7 Hz),3.80 (3H, s), 3.80 (1H, t, J=9.0 Hz), 3.68 (1H, dd, J=3.0, 9.0 Hz), 2.72(1H, d, J=9.0 Hz), 1.67 (3H, s), 1.28 (3H, s). ¹³C NMR (75 MHz, CDCl₃):δ 175.0, 159.2, 134.5, 130.0, 129.2, 129.2, 125.0, 113.7, 113.7, 82.2,73.0, 66.3, 65.4, 61.4, 55.2, 48.5, 45.6, 40.9, 36.6, 29.8, 24.0, 17.1,16.8. HR-MS (ESI) Calcd for [C₂₃H₃₀O₅+H]⁺ 387.2172. found 387.2186.

(1aR,7aS,8R,10aS,Z)-8-(Hydroxymethyl)-1a,5-dimethyl-2,3,6,7,7a,8,10a,10b-octahydrooxi-reno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one,JD209

A mixture of the PMB ether JD208 (44 mg, 0.114 mmol) and2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ, 39 mg, 0.171 mmol) indichloromethane (1.0 mL) and water (0.05 mL) was stirred for 2 h at 0°C. The mixture was diluted with dichloromethane (20 mL) and dried overMgSO₄. The dichloromethane was removed by rotary evaporation and theresidue was purified by flash column chromatography (eluent:hexane/ethyl acetate=1/1) to obtain the desired product, JD209. ¹H NMR(300 MHz, CDCl₃): δ 5.19 (1H, d, J=11.7 Hz), 4.08 (2H, m), 3.87 (1H, t,J=8.7 Hz), 3.76 (1H, dd, J=4.5, 11.7 Hz), 2.75 (1H, d, J=9.0 Hz), 1.67(3H, s), 1.28 (3H, s). ¹³C NMR (75 MHz, CDCl₃): δ 176.4, 134.4, 125.2,82.7, 66.2, 61.6, 58.7, 50.0, 44.8, 40.9, 36.5, 29.8, 24.0, 17.1, 16.8.HR-MS (ESI) Calcd for [C₁₅H₂₂O₄+H]⁺267.1596. found 267.1608.

(1aR,4Z,7aS,8Z,10aS)-1a,5-Dimethyl-8-(piperidin-1-ylmethylene)-2,3,6,7,7a,8,10a,10b-octahydrooxireno[2′,3′:9,10]cyclodeca[1,2-b]furan-9(1aH)-one,JD205

A mixture of the hydroxymethyl analogue, JD209 (20 mg, 0.075 mmol), andDess-Martin periodinane (39 mg, 0.09 mmol) in anhydrous dichloromethane(1.0 mL) was stirred for 2 h at 0° C. The mixture was diluted withdichloromethane (20 mL) and dried over MgSO₄. The dichloromethane wasremoved by rotary evaporation. The crude residue was dissolved inanhydrous methanol (0.5 mL), piperidine (75 4, 0.76 mmol) was added, andthe mixture was stirred at room temperature overnight. The reactionmixture was extracted with ethyl acetate (3×15 mL), washed with waterand brine, and dried over MgSO₄. After removal of the solvent by rotaryevaporation, the crude product was purified by flash columnchromatography (eluent: dichloromethane/MeOH=30/1) to obtain the desiredproduct, JD205. ¹H NMR (300 MHz, CDCl₃): δ 7.17 (1H, d, J=1.8 Hz), 5.21(1H, d, J=12.3 Hz), 3.78 (1H, t, J=6.6 Hz), 1.65 (3H, s), 1.24 (3H, s).¹³C NMR (75 MHz, CDCl₃): δ 174.3, 147.1, 134.3, 125.1, 91.9, 82.1, 66.8,61.3, 51.6, 51.6, 46.9, 40.8, 36.1, 33.3, 26.0, 26.0, 24.4, 23.8, 17.5,17.5. HR-MS (ESI) Calcd for [C₂₀H₂₉NO₃+H]⁺ 332.2226. found 332.2232.

K. REFERENCES

1. Mi J, X Zhang, Z Rabbani, Y Liu, S Reddy, Z Su, F Salahuddin, KViles, P Giangrande, M Dewhirst, B Sullenger, C Kontos, B Clary (2008).RNA aptamer-targeted inhibition of NF-κB suppresses non-small cell lungcancer resistance to doxorubicin. Molecular Therapy 16: 66-73. 2.Al-Hajj M, Wicha M, Benito-Hernandez A, et al. (2003). Prospectiveidentification of tumorigenic breast cancer cells. Proc Natl Acad SciUSA 100: 3983-3988. 3. Hermann P, S Huber, T Herrler, et al. (2007).Distinct populations of cancer stem cells determine tumor growth andmetastatic activity in human pancreatic cancer. Cell Stem Cell 1:313-323. 4. Fillmore C, Kuperwasser C (2008). Human breast cancer celllines contain stem-like cells that self-renew, give rise tophenotypically diverse progeny and survive chemotherapy. Breast CancerRes 10: R25. 5. Ginestier C, Hur M, Charafe-Jauffret E, et al. (2007).ALDH1 is a marker of normal and malignant human mammary stem cells and apredictor of poor clinical outcome. Cell Stem Cell 1: 555-567. 6. LevinaV, Marrangoni A M, DeMarco R, Gorelik E, Lokshin A E (2008).Drug-selected human lung cancer stem cells: cytokine network,tumorigenic and metastatic properties. PLoS One 3:e3077. 7. Jiang F, QQiu, A Khanna, N Todd, J Deepak, L Xing, H Wang, Z Liu, Y Su, S Stass, RKatz (2009). Aldehyde dehydrogenase-1 is a tumor stem cell-associatedmarker in lung cancer. Mol Cancer Res 7: 330-338. 8. Moreb J, H Baker,L-J Chang, M Amaya, M C Lopez, B Ostmark, W Chou (2008). ALDH isozymesdown-regulation affects cell growth, cell motility and gene expressionin lung cancer cells. Mol Cancer 7: 87-105. 9. Zhang D, Qui L, Jin X,Guo Z, Gio C (2009). Nuclear factor-kappaB inhibition by parthenolidepotentiates the efficacy of taxol in non-small cell lung cancer in vitroand in vivo. Mol Cancer Res 7: 1139-1149. 10. Eramo A, F Lotti, G Sette,E Pilozzi, M Biffoni, A Di Virgilio, C Conticello, L Ruco, C Peschle, RDe Maria (2008). Identification and expansion of the tumorigenic lungcancer stem cell population. Cell Death and Differentiation 15: 504-514.11. Marquez-Garban D, Erin L. Maresh, Mohammad Alavi, Hsiao-Wang Chen,Vei Mah, Lora Bagryanova, Steve Horvath, David Chia, Edward Garon, LeeGoodglick, Richard J. Pietras (2011). Progesterone and estrogen receptorexpression and activity in human non-small cell lung cancer. Steroids(in press). 12. Chlebowski R T. Menopausal hormone therapy, hormonereceptor status, and lung cancer in women. Semin Oncol. 2009,36(6):566-71. 13. Wu W, A Onn, T Isobe, S Itasaka et al. (2007).Targeted therapy of orthotopic human lung cancer by combined vascularendothelial growth factor and epidermal growth factor receptor signalingblockade. Molecular Cancer Therapeutics 6: 471-483. 14. Pegram M,Konecny G, O'Callaghan C et al. (2004). Rational combinations oftrastuzumab with chemotherapeutic drugs used in the treatment of breastcancer. J Natl Cancer Inst 96: 739-749. 15. Chou T C, Talalay P (1984).Quantitative analysis of dose-effect relationships: the combined effectsof multiple drugs or enzyme inhibitors. Adv Enzyme Regul 22:27-55 16.C-M Tsai C-M, K-T Chang, L-H Wu, J-Y Chen, A Gazdar, T Mitsudomi, M-HChen, R-P Perng (1996). Correlations between intrinsic chemoresistanceand HER-2/neu gene expression, p53 gene mutations, and cellproliferation characteristics in non-small cell lung cancer cell lines.Cancer Research 56: 206-209. 17. American Cancer Society: Cancer Factsand FIGS. 2012. Atlanta, Ga.: American Cancer Society, 2012 18.Shanmugan R, Kusumanchi P, Appaiah H, Cheng L, Crooks P, Neelakantan S,Peat T, Klaunig J, Matthews W, Nakshatri H, Sweeney C J. A water solubleparthenolide analog suppresses in vivo tumor growth of twotobacco-associated cancers, lung and bladder cancer, by targeting NF-κBand generating reactive oxygen species. Int J Cancer. 201,128(10):2481-94. 19. Zhang D, Qiu L, Jin X, Guo Z, Guo C. Nuclear factorkappaB inhibition by parthenolide potentiates the efficacy of Taxol innon-small cell lung cancer in vitro and in vivo. Mol Cancer Res. 2009,7(7):1139-49. 20. Neelakantan S, Nasim S, Guzman M L, Jordan C T, CrooksP A. Aminoparthenolides as novel anti-leukemic agents: discovery of theNF-kappaB inhibitor, DMAPT (LC-1). Bioorg Med Chem Lett 2009; 19:4346-9.21. Guzman M L, Rossi R M, Neelakantan S, Li X, Corbett C A, Hassane DC, Becker M W, Bennett J M, Sullivan E, Lachowicz J L, Vaughan A,Sweeney C J, et al. (2007). An orally bioavailable parthenolide analogselectively eradicates acute myelogenous leukemia stem and progenitorcells. Blood; 110: 4427-35. 22. Hehner S P, Heinrich M, Bork P M, VogtM, Ratter F, Lehmann V, Schulze-Osthoff K, Droge W, Schmitz M L. (1998).Sesquiterpene lactones specifically inhibit activation of NF-kappa B bypreventing the degradation of I kappa B-alpha and I kappa B-beta. J BiolChem; 273:1288-97. 23. Wen J, You K R, Lee S Y, Song C H, Kim D G.(2002). Oxidative stress-mediated apoptosis. The anticancer effect ofthe sesquiterpene lactone parthenolide. J Biol Chem; 277: 38954-64. 24.Curry E A, III, Murry D J, Yoder C, Fife K, Armstrong V, Nakshatri H,O'Connell M, Sweeney C J. (2004). Phase I dose escalation trial offeverfew with standardized doses of parthenolide in patients withcancer. Invest New Drugs; 22:299-305. 25. Shanmugam R, Jayaprakasan V,Gokmen-Polar Y, Kelich S, Miller K D, Yip-Schneider M, Cheng L,Bhat-Nakshatri P, Sledge G W, Jr, Nakshatri H, Zheng Q-H, Miller M A, etal. (2006). Restoring chemotherapy and hormone therapy sensitivity byparthenolide in a xenograft hormone refractory prostate cancer model.Prostate; 66:1498-511. 26. Nasim S, Crooks P A. Antileukemic activity ofaminoparthenolide analogs. Bioorg Med Chem Lett. 2008; 18(14):3870-3.27. Harris J., M. Lippman, U. Veronesi & W. Willett (1992). Breastcancer. N. Engl. J. Med., 327: 473-451. 28. Pietras, R. J., Arboleda,J., Wongvipat, N., Ramos, L., Parker, M. G., Sliwkowski, M. X., andSlamon, D. J. (1995). HER-2 tyrosine kinase pathway targets estrogenreceptor and promotes hormone-independent growth in human breast cancercells. Oncogene, 10: 2435-2446. 29. Marquez, D, Pietras, R J (2001).Membrane-associated binding sites for estrogen contribute to growthregulation in human breast cancer cells. Oncogene, 20: 5420-5430. 30.Bange J, Zwick E, Ullrich A (2001). Molecular targets for breast cancertherapy and prevention. Nat Med 7: 548-52. 31. Zwick E, Bange J, UllrichA (2001). Receptor tyrosine kinase signaling as a target for cancerintervention strategies. Endocrine-Related Cancer 8: 161-173. 32. SlamonD. J., G. M. Clark, S. G. Wong et al. (1987). Human breast cancer:Correlation of relapse and survival with amplification of the HER-2/neuoncogene. Science, 235: 177-181. 33. Slamon D. J., W. Godolphin, L. A.Jones, J. A. Holt, S. G. Wong, D. E. Keith, W. J. Levin, S. G. Stuart,J. Udove, A. Ullrich and M. F. Press (1989). Studies of the HER-2/neuproto-oncogene in human breast and ovarian cancer. Science, 244:707-711. 34. Wright C., B. Angus, S. Nicholson et al. (1989). Expressionof c-erbB-2 oncoprotein: a prognostic indicator in human breast cancer.Cancer Res., 49: 2087-2094. 35. Nicholson S., C. Wright, J. R. C.Sainsbury, P. Halcrow, P. Kelly, B. Angus, J. R. Farndon and A. L.Harris (1990). Epidermal growth factor receptor as a marker for poorprognosis in node-negative breast cancer patients: neu and tamoxifenfailure. J. Steroid Biochem., 37: 811-818. 36. Benz C., G. Scott, J.Sarup, R. Johnson, D. Tripathy, E. Coronado, H. Shepard and C. Osborne(1993). Estrogen-dependent, tamoxifen-resistant tumorigenic growth ofMCF-7 cells transfected with HER2/neu. Breast Cancer Res. Treatment, 24:85-92. 37. Pegram, M., G. Pauletti and D. Slamon (1998). Her-2/neu as apredictive marker of response to breast cancer therapy. Brst. CancerRes. Trtmt., 52: 65-77. 38. Lipton A, Ali S M, Leitzel K, Demers L,Chinchilli V, Engle L, Harvey H A, Brady C, Nalin C M, Dugan M, CarneyW, Allard J (2002). Elevated serum Her-2/neu level predicts decreasedresponse to hormone therapy in metastatic breast cancer. J Clin Oncol.20:1467-72. 39. Green, S. and P. Chambon (1988). Nuclear receptorsenhance our understanding of transcription regulation. Trends Genet., 4:309-314. 40. Gruber C J, W Tschugguel, C Schneeberger and JC Huber(2002). Mechanisms of disease: Production and actions of estrogens. NewEngl J Med 346: 340-352. 41. Stoica G, Franke T, Wellstein A, CzubaykoF, List H-J, Reiter R, Morgan E, Martin M, Stoica A (2003). Estradiolrapidly activates Akt via the erbB2 signaling pathway. Mol. Endocrinol.17:818-830. 42. Katzenellenbogen B S (1996). Estrogen receptors:bioactivities and interactions with cell signaling pathways. BiolReprod. 54: 287-93. 43. Wakeling A, Nicholson R, Gee J (2001). Prospectsfor combining hormonal and nonhormonal growth factor inhibition. Clin.Cancer Res. 7: 4350s-4355s. 44. Pietras, R. and Szego C. (1977).Specific binding sites for oestrogen at the outer surfaces of isolatedendometrial cells. Nature, 265:69-72. 45. Levin E (1999). Cellularfunctions of the plasma membrane estrogen receptor. Trends Endocrinol.Metabol. 10: 374-377. 46. Marquez D, Pietras R J (2003).Membrane-associated estrogen receptors and breast cancer. In: “TheIdentities of Membrane Steroid Receptors” (Watson C S, editor), KluwerAcademic Publ.: pp. 1-10. 47. Simons K, van Meer G. (1988). Lipidsorting in epithelial cells. Biochemistry 27: 6197-202. 48. Anderson R G(1998). The caveolae membrane system. Annu Rev Biochem 67:199-225. 49.Song K S, Shengwen L, Okamoto T et al. (1996). Co-purification anddirect interaction of ras with caveolin, an integral membrane protein ofcaveolae microdomains. J. Biol. Chem. 271: 9690-9697. 50. Koleske A J,Baltimore D, Lisanti M P (1995). Reduction of caveolin and caveolae inoncogenically transformed cells. Proc Natl Acad Sci USA 92: 1381-5. 51.Pietras R. J. and C. M. Szego (1984). Specific internalization ofestrogen and binding to nuclear matrix in isolated uterine cells.Biochem. Biophys. Res. Commun.,123: 84-90. 52. Chambliss K, Yuhanna I,Mineo C, et al. (2000). Estrogen receptor alpha and endothelial nitricoxide synthase are organized into a functional signaling module incaveolae. Circ Res. 87: E44-E52. 53. Font de Mora J, Brown M (2000).AIB1 is a conduit for kinase-mediated growth factor signaling to theestrogen receptor. Mol Cell Biol 20: 5041-7. 54. Katzenellenbogen B S,Montano M M, Ekena K et al. (1997). Antiestrogens: mechanisms of actionand resistance in breast cancer. Breast Cancer Res Treat 44: 23-38. 55.Konecny G, Pauletti G, Pegram M, et al. (2003). Quantitative associationbetween HER-2 and steroid hormone receptors in hormone receptor-positiveprimary breast cancer. J Natl Cancer Inst. 95:142-153. 56. Wright C., S.Nicholson, B. Angus, J. R. Sainsbury, J. Farndon, J. Cairns, A. L.Harris and C. H. Horne (1992). Relationship between c-erbB-2 proteinproduct expression and response to endocrine therapy in advanced breastcancer. Br. J. Cancer, 65: 118-124. 57. Borg A., B. Baldetorp, M. Ferno,et al. (1994). ErbB2 amplification is associated with tamoxifenresistance in steroid-receptor positive breast cancer. Cancer Letters,81:137-143. 58. Leitzel K., Y. Teramoto, K. Konrad, et al. (1995).Elevated serum c-erbB-2 antigen levels and decreased response to hormonetherapy of breast cancer. J. Clin. Oncol., 13: 1129-1135. 59. DeLaurentis M, Arpino G, Massarelli E, et al. (2000). A meta-analysis ofthe interaction between HER2 and the response to endocrine therapy inmetastatic breast cancer. Am Soc Clin Oncol 19: 301. 60. Hu J C, MokbelK (2001). Does c-erbB2/HER2 overexpression predict adjuvant tamoxifenfailure in patients with early breast cancer? Eur J Surg Oncol 27:335-7. 61. Pietras, R J, Marquez D, Chen H-W, Ayala R, Ramos L, andSlamon D (2003). Improved antitumor therapy with Herceptin and Faslodexfor dual targeting of HER-2 and estrogen receptor signaling pathways inhuman breast cancers with overexpression of HER-2/neu gene. BreastCancer Res Trtmt 82, Suppl 1: 12-13. 62. Kalaitzidis D, Gilmore T D:Transcription factor cross-talk: the estrogen receptor and NF-kappaB.Trends Endocrinol Metab 2005, 16:46-52) 63. Biswas D K, Singh S, Shi Q,Pardee A B, Iglehart J D: Crossroads of estrogen receptor and NF-kappaBsignaling. Sci STKE 2005, 288:pe27 64. Knowlden J, Hutcheson I, Jones Het al. (2003). Elevated levels of EGFR/c-erbB2 heterodimers mediate anautocrine growth regulatory pathway in tamoxifen-resistant MCF-7 cells.Endocrinology 144:1032-44. 65. Wu Y, C Ginther, J Kim, N Mosher, SChung, D Slamon, and J Vadgama (2012). Expression of Wnt3 ActivatesWnt/b-Catenin Pathway and Promotes EMT-like Phenotype inTrastuzumab-Resistant HER2-Overexpressing Breast Cancer Cells. MolCancer Res 2012; 10:1597-1606. 66. Massarweh S, Osborne C K, Creighton Cet al. (2008). Tamoxifen resistance in breast tumors is driven by growthfactor receptor signaling with repression of classic ER genomicfunction. Cancer Res 68:826-33. 67. Marquez D C, Chen H W, Welshons W Vand Pietras R J (2006). Estrogen receptor in membrane lipid rafts andsignal transduction in breast cancer. Molecular Cellular Endocrinol 246:91-100. 68. Berstein L M, Yue W, Wang J P, Santen R J (2011). Isolatedand combined action of tamoxifen and metformin in wild-type,tamoxifen-resistant, and estrogen-deprived MCF-7 cells. Breast CancerRes Treat. 128:109-17. 69. Zhou Y, Yau C, Gray J W, Chew K, Dairkee S H,Moore D H, Eppenberger U, Eppenberger-Castori S, Benz C C (2007).Enhanced NF kappa B and AP-1 transcriptional activity associated withantiestrogen resistant breast cancer. BMC Cancer 7:59. 70. Malik F,Korkaya H, Clouthier S G, Wicha M S (2012). Lin28 and HER2: two stemcell regulators conspire to drive aggressive breast cancer. Cell Cycle11:2780-1. 71. Darling T N, Pacheco-Rodriguez G, Gorio A, Lesma E,Walker C, Moss J (2010). Lymphangioleiomyomatosis and TSC2−/− cells.Lymphat Res Biol. 8(1):59-69. 72. McCormack F X, Inoue Y, Moss J, SingerL G, Strange C, Nakata K, Barker A F, Chapman J T, Brantly M L, Stocks JM, Brown K K, Lynch J P 3rd, Goldberg H J, Young L R, Kinder B W, DowneyG P, Sullivan E J, Colby T V, McKay R T, Cohen M M, Korbee L,Taveira-DaSilva A M, Lee H S, Krischer J P, Trapnell B C; NationalInstitutes of Health Rare Lung Diseases Consortium; MILES Trial Group.Efficacy and safety of sirolimus in lymphangioleiomyomatosis (2011). NEngl J Med 364(17):1595-606. Epub 2011 Mar. 16. 73. Darling T N,Pacheco-Rodriguez G, Gorio A, Lesma E, Walker C, Moss J (2010).Lymphangioleiomyomatosis and TSC2−/− cells. Lymphat Res Biol.8(1):59-69. 74. Astrinidis A, Timothy P Cash, Deborah S Hunter, Cheryl LWalker, Jonathan Chernoff and Elizabeth P Henske (2002). Tuberin, thetuberous sclerosis complex 2 tumor suppressor gene product, regulatesRho activation, cell adhesion and migration. Oncogene 21: 8470-8476. 75.Howe S R, Gottardis M M, Everitt J I, Goldsworthy T L, Wolf D C, WalkerC (1995). Rodent model of reproductive tract leiomyomata: establishmentand characterization of tumor-derived cell lines. Am J Pathol146:1568-79. 76. Yu J J, Robb V A, Morrison T A, Ariazi E A,Karbowniczek M, Astrinidis A, Wang C, Hernandez-Cuebas L, Seeholzer L F,Nicolas E, Hensley H, Jordan V C, Walker C L, Henske E P (2009).Estrogen promotes the survival and pulmonary metastasis of tuberin-nullcells. Proc Natl Acad Sci USA 106(8):2635-40. Epub 2009 Feb. 6. 77.Arbiser J L, Yeung R, Weiss S W, Arbiser Z K, Amin M B, Cohen C, FrankD, Mahajan S, Herron G S, Yang J, Onda H, Zhang H B, Bai X, Uhlmann E,Loehr A, Northrup H, Au P, Davis I, Fisher D E, and Gutmann D H. Thegeneration and characterization of a cell line derived from a sporadicrenal angiomyolipoma: use of telomerase to obtain stable populations ofcells from benign neoplasms. Am J Pathol 159: 483-491, 2001. 78. Yu J,Astrinidis A, Howard S, Henske E P. Estradiol and tamoxifen stimulateLAM-associated angiomyolipoma cell growth and activate both genomic andnongenomic signaling pathways. Am J Physiol Lung Cell Mol Physiol. 2004April; 286(4):L694-700. Epub 2003 Aug. 15.

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated by reference in theirentirety for all purposes.

What is claimed is:
 1. A compound, or a pharmaceutically acceptable salt thereof, wherein the compound has a formula selected from the group consisting of:

wherein, L⁰ is independently a bond or an unsubstituted C₁-C₁₀ alkylene; and R¹ and R² are independently hydrogen, —OH, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; where R¹ and R² may optionally be joined to form a substituted or unsubstituted heterocycloalkyl or a substituted or unsubstituted heteroaryl; with the proviso that the compound, or the pharmaceutically acceptable salt thereof is not


2. The compound, or a pharmaceutically acceptable salt thereof, of claim 1, wherein the compound has a formula selected from the group consisting of:


3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R¹ is L¹-R³; L¹ is independently a bond or an unsubstituted C₁-C₁₀ alkylene; and R³ is independently hydrogen, halogen, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
 4. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein R³ is independently hydrogen, halogen, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂, 4-methoxybenzyl, 4-methoxyphenyl, or pyridyl.
 5. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R² is L²-R⁴; L² is independently a bond or an unsubstituted C₁-C₁₀ alkylene; and R⁴ is independently hydrogen, halogen, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.
 6. The compound of claim 5, or a pharmaceutically acceptable salt thereof, wherein R⁴ is independently hydrogen, halogen, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH₂CH₃, —CH₂CH₂OH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂, 4-methoxybenzyl, 4-methoxyphenyl, or pyridyl.
 7. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R¹ and R² are joined to form an unsubstituted heterocycloalkyl.
 8. The compound, or a pharmaceutically acceptable salt thereof, of claim 1, wherein the compound is selected from the group consisting of


9. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound, or pharmaceutically acceptable salt thereof, of claim
 1. 10. The pharmaceutical composition of claim 9, comprising a second agent wherein said second agent is an anti-cancer agent.
 11. A method of treating cancer in a patient in need of said treatment, said method comprising administering a therapeutically effective amount of a compound of claim 1, or a pharmaceutically acceptable salt thereof, to said patient.
 12. The method of claim 11, wherein said cancer is breast cancer.
 13. The method of claim 12, wherein said breast cancer is resistant to an anti-cancer agent.
 14. The method of claim 11, wherein said cancer is lung cancer.
 15. The method of claim 14, wherein said lung cancer is resistant to an anti-cancer agent.
 16. A method of treating a non-malignant hyperproliferative disease in a patient in need of said treatment, said method comprising administering a therapeutically effective amount of a compound of claim 1, or a pharmaceutically acceptable salt thereof, to said patient.
 17. A method of inhibiting cancer cell growth or survival comprising contacting the cell with an effective amount of a compound of claim 1, or a pharmaceutically acceptable salt thereof.
 18. The method of claim 17, wherein said cancer cell is a cancer stem cell.
 19. A method of decreasing the level of activity of NF-κB or mTOR in a cell comprising contacting the cell with an effective amount of a compound of claim 1, or a pharmaceutically acceptable salt thereof.
 20. A method of increasing the level of activity of TSC1 or TSC2 in a cell comprising contacting the cell with an effective amount of a compound of claim 1, or a pharmaceutically acceptable salt thereof. 